Neurobiology Research Reagents Research

Neurobiology Markers 1-4

Alzheimers disease 4-9

Parkinsons disease 9-10

Brain tumours 10

Neuromuscular disease 11

Neuropathic pain 12

...Neurobiology ... Neurochemistry ... Neuropathology ...Neurobiology ... Neurochemistry ... Neuropathology ...Neurobiology ... Neurochemistry ... Neuro Neurobiology Markers

Neurobiology is the foundation of multiple fields of research, from developmental biology to neurological pathology. The cells of the nervous system and their organisation into functional pathways, serve to process information and regulate behaviour. The nervous system comprises both neurons and glia cells. Neurons receive, propagate and transmit electrochemical

www.alphalabs.co.uk impulses whilst glia provide mechanical and metabolic support. Microglia

Microglia are the resident macrophages of the brain and act as the main form of active immune defence scavenging damaged neurons, debris and pathogens. Whilst microglia offer a protective function they can also have deleterious effects as a result of cytotoxic secretion of hydrogen peroxide, nitric oxide, proteases and cytokines.

Microglial cells are the only brain cells to express Iba-1 (ionised calcium binding adapter molecule 1). Calcium ions are important signal mediators and exert their signalling activity through association with various calcium binding proteins. Iba-1 is a 17kDa protein from the large EF hand family of proteins which contain the EF-hand motif. Iba-1 expression is up-regulated in activated microglia enabling differentiation between cells engaged in routine surveillance and those which are activated in response to injury. For this reason Iba-1, also known as Allograft Inflammatory factor 1 (AIF-1) is often used in immunohistochemistry as a marker for microglia. Enhanced Iba-1 expression has been observed in traumatic brain injury, ischemia and inflammation.

The Wako Anti-Iba-1 antibodies for immunocytochemistry and Western Blotting have been raised against a synthetic peptide corresponding to the carboxyl-terminus of Iba-1. This is conserved amongst human, rat and mouse Iba-1 protein sequences. These antibodies are specific to microglia and macrophages and do not cross react with neurons or astrocytes.

The immunocytochemistry Iba-1 antibody is well suited for double- immunostaining of brain tissue or cell cultures in combination with a

monoclonal antibody specific to astrocytes (Glial Fibrillary Acid Protein (GFAP)) as seen opposite. Above: Immuno-double staining of rat primary mixed cell culture:

Iba-1 Green: Iba1 (Wako Cat. 019-19741) Cat. No. Description Pack Size Red: Astrocyte, which reacts to anti-GFAP, mAb 019-19741 Anti-IBA1 Rabbit IC 50µg (Data was provided by Dept. of Neurochemistry, 016-20001 Anti-IBA1 Rabbit WB 50µg National Institute of Neuroscience (Japan)) Not for diagnostc use

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2 Olfactory Sensory Neurons

Olfactory neurons extend dendrites to the surface of the olfactory epithelium lining the nasal cavity and project unmyelinated axons back into the olfactory bulb situated above the nasal cavity and below the frontal lobe. When odorant molecules bind to specific sites on olfactory receptors, either by diffusion or interaction with odorant binding molecules, this results in

coupling of the receptor to a G protein to stimulate adenylyl cyclase. Rising www.alphalabs.co.uk cAMP opens a cyclic nucleotide-gated ion channel and the resulting influx of ions instigates depolarisation which in turn triggers an action potential.

Olfactory Marker Protein (OMP) is a 19KDa cytosolic protein expressed Above: Immunofluorescence staining of adult predominantly by mature, functioning, olfactory neurons. It is not mouse olfactory epithelium with goat anti-OMP expressed in neural precursor basal cells. OMP has long been recognised as visualised with Cy2 (Jackson Immuno Research). a molecular marker for olfactory neurons. The Wako Anti-Olfactory Marker (Data was provided by Dr Frabk L Marglois and Protein anti-sera has been raised against rodent OMP and is highly specific Dr Jae Hyung Koo, Department of Anatomy and for mature olfactory neurons, their axons and terminals in rodents humans, Neurobiology, School of Medicine, University of Maryland.) marsupials and amphibians. This anti-sera is suitable for Western Blot and immunocytochemistry applications.

Olfactory Marker Protein Cat. No. Description Pack Size 544-10001 Anti-Olfactory Marker Protein 100µl Acetylcholinesterase

Acetylcholinesterase (AChE) terminates synaptic transmission by degrading Acetylcholine (ACh) and enabling the neuron to return to its resting state. Inhibitors of AChE can increase the concentration of ACh in the synaptic cleft and impede neurotransmission. Irreversible AChE inhibitors such as organophosphates can lead to muscular paralysis, convulsions and asphyxiation. Reversible cholinesterase inhibitors are used therapeutically and have been used in the treatment of Alzheimer’s disease, dementia with Lewy bodies and myasthenia gravis. The BioPorto mouse monoclonal AChE antibodies for human and bovine brain AChE are suitable for ELISA and Immunoblotting applications.

Acetycholinestersase Antibodies

Cat. No. Description Pack Size HYB 101-03-02 Anti-Acetylcholinesterase (AChE) MAb 200µg HYB 101-03-1 Anti-Acetylcholinesterase (AChE) MAb 1mg HYB 111-05-02 Anti-Acetylcholinesterase (AChE) MAb 200µg HYB 111-05-1 Anti-Acetylcholinesterase (AChE) MAb 1mg HYB 190-01-02 Anti-Acetylcholinesterase (AChE) MAb 200µg HYB 190-01-1 Anti-Acetylcholinesterase (AChE) MAb 1mg Not for diagnostc use

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3 Synaptic Receptors

Neurotransmission is the process by which neurotransmitters released by one neuron activate the receptors of a second neuron. The neurotransmitter is released by the pre-synaptic neuron and crosses the synaptic cleft to activate a receptor on the post synaptic neuron. Neurotransmitters are stored in membrane-bound vesicles at the pre-synaptic membrane and

www.alphalabs.co.uk their release is initiated by depolarisation of the neuron followed activation of voltage-gated calcium channels. The calcium influx triggers fusion of synaptic vesicles with the plasma protein in a process mediated by SNARE (soluble N-ethylmalemide-sensitive factor attachment protein receptor) proteins.

The SNARE proteins syntaxin and SNAP-25 are found in the plasma membrane whilst a third SNARE protein synaptobrevin is found in the vesicle membrane. The assembly of the trans-SNARE complex is thought to bridge the opposing lipid layers to enable fusion.

The Wako Syntaxin and SNAP-25 monoclonal antibodies for immunochemistry were raised against rat synaptosome and are suitable for Western Blot and indirect immunofluorescence applications.

Synaptic Receptors Cat. No. Description Pack Size 014-15771 Anti-Rat SNAP-25 MAb 100µg 011-15781 Anti-Rat Syntaxin MAb 100µg Alzheimer’s disease

Alzheimer’s disease is the most common cause of dementia. During the progression of the disease, protein plaques and neurofibrilllary tangles develop in the brain. The senile plaques that form are dense, insoluble deposits of amyloid-β peptide (Aβ) and other materials which accumulate outside and around neurons.

Aβ peptide can exist in both 40 and 42 amino acid forms. Aβ42 aggregates more readily than Aβ40 and it appears that Aβ42 is deposited initially.

Early stage, diffuse plaques may be positive for Aβ42 but negative for Aβ40. Plaques containing Aβ40 may only appear in the more advanced stages of disease.

Post-translational modifications of Aβ such as truncation and pyroglutamate formation, which typically effect the N-terminus, have been reported. This may influence protein properties such as solubility and aggregation. Not for diagnostc use

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4 Wako offers an extensive range of ELISA assay kits for detection of Aβ ■ BAN50: Detects the N-terminal of Aβ peptides. These versatile ELISA kits can be used to analyse various human ■ BNT77: Detects Aβ(11-28) and rat samples including plasma, cerebrospinal fluid, tissue homogenate ■ BA27: Detects the C-terminal of Aβ(40) or cell culture media. Wako ELISA kits utilise monoclonal antibodies ■ BC05: Detects the C-terminal of Aβ(42) developed by Takeda Chemical Industries Ltd, the assay range includes kits for detection of both Aβ 42 and Aβ40, with both full-peptide and N-terminal variants. www.alphalabs.co.uk Using the BAN50 antibody as capture and biotinylated BA27 for detection, the human Aβ (1-40) ELISAs target the whole protein as the antibody combination recognises both the N- and C-terminus of Aβ40 peptide(1-40). In addition to the original assay (292-62301) a second

assay has been developed using the (Fab’)2 fragment of the labelled BA27 to achieve greater antigen-antibody stability (298-64601).

Similarly the BAN50/BC05 antibody combination targets the whole Aβ42 peptide (1-42) and both standard range and high-sensitivity kits are available.

The human/rat Aβ40 and Aβ42 ELISAs use the BNT77 antibody for detection which binds Aβ(11-28) and will therefore detect both complete and N-terminus modified proteins, Aβ(x-40) and Aβ(x-42) respectively.

Cat. no. Antibody pairing Range βAmyloid, measured (pmol/L) human human rat (mouse) rat (mouse) Aβ(x-40) Aβ(x-42) Aβ(1-40) Aβ(1-42) Aβ(1-40) Aβ(1-42) 292-62301 BAN50/BA27(Fab’) 1.0-100 • - - - - -

298-64601 BAN50/BA27(Fab’)2 1.0-100 • - - - - - 298-62401 BAN50/BC05(Fab’) 1.0-100 - • - - - - 296-64401 BAN50/BC05 (Fab’) 0.1-20.0 - • - - - - 294-62501 BNT77/BA27(Fab’) 1.0-100 • - • - • -

294-64701 BNT77/BA27(Fab’)2 1.0-100 • - • - • - 290-62601 BNT77/BC05(Fab’) 1.0-100 - • - • - • 292-64501 BNT77/BC05(Fab’) 0.1-20.0 - • - • - •

Human Amyloid β ELISA Cat. No. Description Pack Size 292-62301 Human β Amyloid 1-40 ELISA 96 wells 298-64601 Human β Amyloid 1-40 ELISA II 96 wells 298-62401 Human β Amyloid 1-42 ELISA 96 wells 296-64401 Human β Amyloid 1-42 ELISA H/S 96 wells

Human/Rat Amyloid β ELISA Cat. No. Description Pack Size 294-62501 Human/Rat β Amyloid 40 ELISA 96 wells 294-64701 Human/Rat β Amyloid 40 ELISA II 96 wells 290-62601 Human/Rat β Amyloid 42 ELISA 96 wells 292-64501 Human/Rat β Amyloid 42 ELISA H/S 96 wells Not for diagnostc use

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5 Amyloid Precursor Protein (APP) is an integral membrane protein with concentrated expression in neuron synapses. It is the precursor of Aβ which accumulates in the brains of Alzheimer’s sufferers. APP is subject to cleavage by the proteolytic secretase enzymes (α-, β- and γ-secretase). The predominant processing pathway for APP is proteolysis by α-secretase, which cleaves APP within the fragment that gives rise to Aβ, hence the α-secretase pathway precludes Aβ formation. β–secretase (also known as β-site APP Cleaving Enzyme 1 or BACE1) cleavage results in a membrane www.alphalabs.co.uk bound fragment known as C99. Subsequent cleavage by γ-secretase produces Aβ. This has led to interest in β-secretase inhibitors which may prevent or slow the build up of Aβ and be of therapeutic use in the management of Alzheimer’s disease.

β- Secretase Cat. No. Description Pack Size Inhibitors 115-00901 KMI-429 1mg 112-00911 KMI-574 1mg 119-00921 KMI-1072 1mg 116-00931 KMI-1303 1mg Substrate 334-32121 MOCAc-SEVNLDAEFRK (Dnp) RR-NH2 1mg

γ- Secretase Cat. No. Description Pack Size Inhibitors 333-43941 L-685,458 1mg 333-32191 (3,5-Difluorophenylacetyl)-Ala-Phg-Obut 5mg Substrate 339-32171 Nma-Gly-Gly-Val-Val-Ile-Ala-Thr-Val-Lys 1mg

Neurofibrillary tangles are the result of protein aggregation within the cell. The neuron cytoskeleton serves to guide molecules between the body of the cell and the end of the axon. The microtubule-associated Tau protein becomes hyperphosphorylated and then aggregates with other threads producing neurofibrillary tangles and disrupting the neuron’s transport system. The BF-168 probe selectively binds plaques in Alzheimer’s disease brain sections and recognises Aβ(1-42) positive diffuse plaques. The second probe, BF-170, binds neurofibrillary tangles exhibiting a greater

binding affinity to Tau fibrils, neurophil threads and paired helical filament than to Aβ fibrils.

(A) Senile plaque staining with BF-168, (B) 6F/3D- Immunofluorescent Probes Anti-Aβ antibody immunostaining (C) Staining of Cat. No. Description Pack Size neurofibrillary change using BF-170 (D) Anti-phos- phor-Tau antibody (pSer422). 029-16361 BF-168 1mg (Data provided by Prof. Kudo Yukitsuka of 026-16371 BF-170 1mg Toohoku Univ. Biomedical Engineering Research

Not for diagnostc use Organisation)

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6 Another fluorescent probe for detection of β amyloid plaques is PP-BTA-1, a newly developed push-pull benzothiazole derivative. PP-BTA-1 consists of benzothiazole as the highly polarised bridge, a dimethylamino group as the donor and a ducyanomethylene group as the acceptor. The PP-BTA-1 probe selectively binds Aβ, exhibiting a stronger binding than the PET ligand [11C]-2-(4-(methylamino) phenyl)-6- hydroxybenzothiazole (PIB). PP-BTA-1 also has less adverse effects on absorption by self-fluorescence or biogenic factors in histological staining, www.alphalabs.co.uk because of the wavelength of this probe is close to the near-infrared region.

Histological staining with PP-BTA-1 has been successfully used in brain tissue sections from both human Alzheimer’s disease and animal models, such as the TG2576 mouse, to investigate Aβ plaques by fluorescent microscopy. PP-BTA-1

■ [[6-(Dimethylamino) benzothiazole-2-yl] methylene] malononitrile ■ Solubility: Soluble in ethanol

■ λex: 540nm; λem: 634nm (in EtOH)

Figure 1.

Figure 1. Neuropathological staining of PP- BTA-1 in 10µm brain section from an animal model of AD, the Tg2576 mouse (A) and wild- type normal mouse (C). Aβ plaques labelled with PP-BTA-1 were confirmed by staining of the serial section using thioflavin (B). Senile plaques on Tg2576 mouse brain section were selectively fluorescent-stained with PP-BTA-1 (A) as well as thioflavin S (B). On the other hand, no parts on wild-type normal mouse brain sections were stained with PP-BTA-1 (C). Figure 2.

Figure 2. Neuropathological staining of 5µm AD brain sections from the temporal cortex (A, B) and adult normal temporal brain section (C). Many neuritic plaques are clearly stained with PP-BTA-1 (A). Intense fluorescence can be seen in the core of neuritic plaques. Aβ immunostaining with anti Aβ anitbodies in the serial secions shows an idential staining pattern of plaques (B). Senile plaques on AD brain section were selectively fluorescent-stained with PP-BTA-1 (A) as well as anti Aβ antibody. On the other hand, PP-BTA-1 no parts on an adult normal temporal brain Cat. No. Description Pack Size section were stained (C).

165-25301 PP-BTA-1 1mg (Data provided by Masahiro Ono and Hideo Saji at School of Pharmaceutical Sciences, Kyoto University) Not for diagnostc use

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7 The neurotransmitter glutamate may also play a role in neurodegenerative disease. Activated microglia release glutamate which can induce excitotoxic neuronal death. N-methyl-D-aspartate (NDMA) receptor signalling increases calcium ion influx and Ca2+/calmodulin-dependant kinase activation (CaMK), in turn activating neuronal nitric oxide synthase (nNOS) which increases nitric oxide production. High concentrations of NO exert neurotoxic effects; shutting down the mitochondrial respiratory chain, impairing dendritic and axonal transport and eventually leading to www.alphalabs.co.uk cell death. Glutamate receptor blockade may offer a potential therapeutic approach to neurodegenerative disease.

Gap junctions, adjacent hemichannels that directly connect the cytoplasmic compartments of adjacent cells, allow free passage of ions and small molecules between cells. Such gap junctions are important in excessive glutamate release and blockade of these hemichannels by glycyrrhetinic acid derivatives may prevent microglia/macrophage-mediated neuronal death. Glycyrrhetinic acid and its derivatives offer poor penetration of the blood- brain barrier (BBB) but INI-0602 is a novel BBB permeable gap junction hemichannel blocker shown to inhibit glutamate mediated toxicity.

NDMA Antagonists Cat. No. Description Pack Size 041-31941 DL-AP7 5mg 031-22091 (R)-CPP 1mg 037-22093 (R)-CPP 5mg 060-05861 Felbamate 10mg 066-05863 Felbamate 50mg 138-16983 Memantine Hydrochloride 100mg 166-25191 Pentamidine Isethionate 50mg 018-04821 D-Aspartic Acid 5g 016-04822 D-Aspartic Acid 25g 013-04832 L-Aspartic Acid 25g 015-04831 L-Aspartic Acid 100g (A) Fluorescent microscopic images of live/ 017-04835 L-Aspartic Acid 500g dead staining of mouse primary cortical neuron 041-32443 6-Diazo-5-oxo-L-norleucine 25mg cultures in microglia-conditioned media. Neurons in microglia-conditioned media NDMA Agonists containing 1 µg/ml LPS (LPS) underwent cell death with neuritic beading, whereas neurons Cat. No. Description Pack Size in microglia-conditioned media containing 174-00271 2,3-Pyridinedicarboxylic Acid 1g PBS (PBS), 1 µg/ml LPS plus 100 µM CBX 172-00272 2,3-Pyridinedicarboxylic Acid 25g (100 µM CBX), or 1 µg/ml LPS plus 100 µM INI-0602 (100 µM INI-0602) appeared healthy.

191-08821 D-Serine 1g Green, neuron (MAP2); red, dead cell (PI); 197-08823 D-Serine 5g blue, nucleus (Hoechst). Scale bar, 20 µm. (B) Glutamate concentrations in media. (C) The 199-08822 D-Serine 25g percentages of dead neurons. Data represent Gap Junction Hemichannel Inhibitor the means ± SD (n = 6 per group). *, P<0.05 vs. PBS; **,P<0.05 vs. LPS; †, P<0.05 vs. 1 µM; Cat. No. Description Pack Size §, P<0.05 vs. 10 µM. 097-06511 INI-0602 1mg Data is referred from Takeuchi, H et al, 2011. 093-06513 INI-0602 5mg

Not for diagnostc use PLoS ONE, 6, e21108

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8 Inflammation is a common feature of neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. Naturally occurring phytochemicals with anti-oxidant properties have been studied in conjunction with such conditions, as these may represent alternative candidates for neurodegenerative disease therapy.

The polyphenols carnosol and rosmarinic acid are components of the herb rosemary, with anti-inflammatory and neuroprotective actions. www.alphalabs.co.uk Similarly the yellow spice turmeric contains three curcuminoids; curcumin, desmethoxycurcumin and bis-desmethoxycurcumin, these anti-oxidant polyphenols have been studied in models of Alzheimer’s and Parkinson’s disease.

Reseveratrol is another anti-oxidant with anti-inflammatory properties, found in the skins of red grapes, it has also demonstrated neuroprotective effects in various animal models of neurodegenerative disease. Such agents have also shown anti-cancer effects.

Phyto-chemicals Cat. No. Description Pack Size 036-22161 Carnosol 1mg 032-22163 Carnosol 5mg 182-02691 Rosmarinic Acid 50mg 188-02693 Rosmarinic Acid 500mg 181-01723 Resveratrol 500mg 185-01721 Resveratrol 100mg 036-04922 Curcumin 25g 038-04921 Curcumin 1g Parkinson's disease

Parkinson’s disease (PD) is a neurodegenerative disorder characterised by cell loss within the substantia nigra and symptoms of bradykinesia, rigidity and tremor. Lewy bodies (LB), aggregations of protein that form within nerve cells, are a hallmark of PD but also occur in dementia with Lewy bodies (DLB).

The ubiquitin-proteasome system removes unwanted proteins that are no longer required by the cell and dysfunction of this system results in a build of up protein within the cell, including α-synuclein which is a major component of LB. Not for diagnostc use

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9 Lewy Bodies (LB) are α-synuclein-immunoreactive inclusions comprising a number of neurofilament proteins together with proteins responsible for proteolysis. One of the first sites for deposition of LB is the olfactory bulb. As the disease progresses areas of the midbrain, basal forebrain and eventually the neocortex are affected.

Whilst the mechanisms of α-synuclein aggregation are still not fully understood, it appears that much of it found in LB is phosphorylated. www.alphalabs.co.uk Phosphorylation may also influence cell toxicity.

The Wako Anti-phosphorylated α-synuclein antibody is ideal for looking at expression of α-synuclein protein phosphorylated at serine 129, in both human and rodent tissue. (A) Temporal neocortex of DLB brains were immunostained with anti-Phosphorylated Suitable for both Western Blot and immunohistochemistry (IHC) α-Synuclein. [Big arrow and mini arrow] applications as seen opposite, this monoclonal antibody is produced indicate LBs and Lewy neutrites respectively. (B) Brainstem LBs in pigmented neurons of the by the hybridoma clone pSyn#64 and does not cross react with un- substantia nigra in PD. (C) Western blot analysis phosphorylated α-synuclein protein. of α-Synuclein differentially extracted with urea from cerebral cortices of a patient with DLB (D) and a normal control (C) Individual Anti-Phosphorylated α-synuclein probed with monoclonal antibody pSyn#64 (Anti-Phosphorylated α-synuclein). This Cat. No. Description Pack Size antibody strongly reacted with the urea-soluble 015-25191 Anti-Phosphorylated Alpha Synuclein 50µl phosphorylated α-synuclein in DLB brains. Astrocytic tumours

Gliomas are tumours that start in the brain or spine with varying malignancy. Astrocytomas are one of the most common forms of primary brain tumour that develop in astrocytes, an abundant subset of glial cells in the nervous system that serve a number of structural and biochemical support roles.

There is a WHO classification for grading astrocytomas with regard to malignancy:

■ Pilocytic astrocytoma (Grade I) ■ Diffuse astrocytoma (Grade II) ■ Anaplastic astrocytoma (Grade III) ■ Glioblastoma (Grade IV).

Frequent mutations of Isocitrate dehydrogenase (IDH) have been reported in grade II/III glioma or in stage IV glioblastoma that has developed from low-grade gliomas. Isocitrate dehydrogenase is an oxidoreductase that interconverts isocitric acid and α-ketoglutarate. Not for diagnostc use

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10 The Wako IDH monoclonal antibodies are suitable for use in ELISA, Western Blot and immunochemistry applications. The IDH1 and IDH2 antibodies are raised against both wild type and mutated isozymes and cross react with human, mouse and hamster. Antibodies are also available to detect the IDH1-R132H and IDH1-R132S mutations.

IDH Antibodies Cat. No. Description Pack Size www.alphalabs.co.uk 014-24061 Anti-IDH1 MAb 100µg 011-24071 Anti-IDH2 MAb 100µg 018-24081 Anti-IDH1-R132H MAb 100µg 015-24091 Anti-IDH1-R132S MAb 100µg

Podoplanin, a type I transmembrane protein also known as Aggrus, gp44 and T1α has also been associated with brain tumours. It has been reported that increased concentrations of podoplanin in brain tumours may correlate with malignancy. Podoplanin has many reported effects and features a platelet aggregation-stimulating (PLAG) domain at the N-terminus. Whilst podoplanin is not expressed in vascular endothelial cells it is present in lymphatic endothelium prompting investigation of podoplanin as a lymph vessel marker. The Wako monoclonal antibodies for human and mouse podoplanin are suitable for ELISA, Western Blot (WB), Immunocytochemistry (ICC), flow cytometry and immunoprecipitation techniques.

Podoplanin Antibodies Cat. No. Description Pack Size 018-24101 Anti-Human Podoplanin 100µg 015-24111 Anti-Mouse Podoplanin 100µg Neuromuscular disorders

Neuromuscular disorders affect muscles and/or their direct nervous control. Muscular dystrophies (MDs) are a group of disorders characterised by progressive skeletal muscle weakness and may involve eye abnormalities; such as congenital MD muscle-eye-brain disease. Pikachurin is a dystroglycan-interacting protein which plays an essential role in photo receptor ribbon synapse and bipolar dendrite interaction. A binding abnormality between pikachurin and dystroglycan is associated

with MDs and is important for retina physiology. This rabbit pAb can be used to detect mouse pikachurin by WB or IHC and may be of use to those investigating synaptic signal transmission and visual function regeneration.

Pikachurin Cat. No. Description Pack Size 011-22631 Anti-Mouse Pikachurin 50µl Not for diagnostc use

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Spinal Muscular Atrophy (SMA) is a lower motor neuron condition causing weakness and muscle wasting and is an autosomal recessive disorder linked to chromosome 5q13. Several genes have been associated with SMA, including Neuronal apoptosis inhibitory protein (NAIP). The deletion/mutation of this gene in some SMAs is thought to contribute to motor neuron dysfunction and cell death.

NAIP expression has also been associated with neurodegenerative diseases such as Alzheimer’s and multiple sclerosis. Anti-Human NAIP rabbit antiserum is suitable for western blotting and detects a band at 140kDa.

Neuronal Apoptosis Inhibitory Protein Cat. No. Description Pack Size 019-24251 Anti-Human NAIP 20µl Neuropathic pain

Purinergic receptors are plasma membrane molecules found in various mammalian tissues. The P2X subset of receptors are ligand-gated ion channels, nonselective cation channels that mediate excitatory postsynaptic responses and are widely expressed in neurons and glial cells throughout the central and peripheral nervous systems. www.alphalabs.co.uk The P2X4 receptor has been linked to neuropathic pain mediated by microglia and increased expression of the receptor is seen following spinal injury. This

mAb detects rat P2X4 receptors and is suitable for WB and ICC applications.

Anti-Rat P2X4 Cat. No. Description Pack Size 016-23281 Anti Rat P2X4 MAb (RUO) 50µg Not for diagnostc use

Neuroimmunology and Autoimmunity

Bühlmann's CE IVD marked ELISA assays are available for detection of autoantibodies to:

■ GM1 ■ Combinations of GA1, GM1, GM2, GD1a, GD1b, GQ1b ■ MAG ■ SGPG

Find out more at: www.alphalabs.co.uk/neuroimmunology Diagnostic Tests

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12 Bibliography

Selected Bibliography

Anti-IbaI Toll-Like Receptor 3 Expression in Glia and Neurons Alters in Response to White Matter Injury in Preterm Infants. Vontell, R., Supramaniam, V., Thornton, C., Wyatt-Ashmead, J., Mallard, C., Gressens, P., Rutherford, M., & Hagberg, H. Dev Neurosci (2013): 35(2-3): 130-139.

Microglial activation, increased TNF and SERT expression in the prefrontal cortex define stress-altered behaviour in mice www.alphalabs.co.uk susceptible to anhedonia. Couch, Y., Anthony, D.C., Dolgov, O., Revischin, A., Festoff, B., Santos, A. I., Steinbusch, H. W., & Strekalova, T. Brain Behav Immun (2013): 29:136-146.

Interleukin-1 Mediates Neuroinflammatory Changes Associated with Diet-Induced Atherosclerosis. Denes, A., Drake, C., Stordy, J., Chamberlain, J., McColl, B.W., Gram, H., Crossman, D., Francis, S., Allan, S.M. & Rothwell, N.J. J Am Heart Assoc. (2012):1(3):e002006

Brain Inflammation is induced by co-morbidities and risk factors for stroke. Drake, C., Boutin, H., Jones, M.S., Denes, A., McColl, B.W., Selvarajah, J.R., Hulme, S., Georgiou, R.F., Hinz, R., Gerhard, A., Vail, A., Prenant, C., Julyan, P., Maroy, R., Brown, G., Smigova, A., Herholz, K., Kassiou, M., Crossman, D., Francis, S., Proctor, S.D., Russell, J.C., Hopkins, S.J., Tyrrell, P.J., Rothwell, N.J. & Allan, S.M. Brain Behav Immun (2011): 25(6): 1113-1122

Selective white matter pathology induces a specific impairment in spatial working memory. Coltman, R., Spain, A., Tsenkina, Y., Fowler, J.H., Smith, J., Scullion, G., Allerhand, M., Scott, F., Kalaria, R.,N., Ihara, M., Daumas, S., Deary, I.J, Wood, E., McCulloch, J., & Horsburgh, K. Neurobiol Aging (2011): 32(12): 2324.e7-12.

Brief, low frequency stimulation of rat peripheral C-fibres evokes prolonged microglial-induced central sensitisation in adults but not in neonates. Hathway, G.J., Vega-Avelaira, D., Moss, A., Ingram, R., & Fitzgerald, M. Pain (2009): 144(1-2):110-118

Macrophage/microglia-specific protein Iba1 enhances membrane ruffling and Rac activation via phospholipase C-gamma-dependent pathway. Kanazawa, H., Ohsawa, K., Sasaki, Y., Kohsaka, S. & Imai, Y. J Biol Chem (2002):277(22):20026-20032.

Involvement of Iba1 in membrane ruffling and phagocytosis of macrophages/microglia. Ohsawa, K., Imai, Y., Kanazawa, H., Sasaki, Y., Kohsaka, S. J Cell Sci (2000):113(17):3073-3084.

Microglia-specific localisation of a novel calcium binding protein, Iba1. Ito, D., Imai, Y., Ohsawa, K., Nakajima, K., Fukuuchi, Y., Kohsaka, S. Brain Res Mol Brain Res (1998): 57(1):1-9.

A novel gene iba1 in the major histocompatibility complex class III region encoding an EF hand protein expressed in a monocytic lineage. Imai, Y., Ibata, I., Ito, D., Ohsawa, K., & Kohsaka, S. Biochem Biophys Res Commun (1996): 224(3):855-62.

13 Bibliography

Anti-OMP Olfactory ensheathing glia are required for embryonic olfactory axon targeting and the migration of gonadotropin- releasing hormone neurons. Barraud, P., St John, J.A., Stolt, C.C., Wegner, M., & Baker, C.V. Biol Open (2013):2(7):750-759.

OMP-ZsGreen fluorescent protein transgenic mice for visualisation of olfactory sensory neurons in vivo and in vitro. Ekberg, J.A., Amava, D., Chehrehasa, F., Lineburg, K., Claxton, C., Windus, L.C., Key, B., Mackay-Sim, A. & St John, J.A. J Neurosci Methods (2011): 196(1): 88-98. www.alphalabs.co.uk

Prion Shedding from Olfactory neurons into nasal secretions. Bessen, R.A., Shearin, H., Martinka, S., Boharski, R., Lowe, D., Wilham, J.M., Caughey, B. & Wiley, J.A. PLoS Pathog (2010): 6(4): e1000837

Bex1 knock out mice show altered skeletal muscle regeneration. Koo, J.H, Smiley, M.A., Lovering, R.M., & Margolis. F.L. Biochem Biophys Res Commun (2007): 363(2):405-410.

The interaction of Bex and OMP reveals a dimer of OMP with a short half-life. Koo, J.H., Gill, S., Pannell, L.K, Menco, B.P., Margolis, J.W. & Margolis, F.L. J Neurochem (2004):90(1):102-116.

Isolation and Characterisation of Rat Olfactory Marker Protein. Keller, A. & Margolis, F.L. J Biol Chem (1976): 251(20):6232-6237.

SNAP-25 & Syntaxin Evidence for the putative docking/fusion complex of exocytosis in parotid acinar cells. Takuma, T., Tagaya, M. & Ichida, T. FEBS Letters (1997):404(1):34-36.

Beta Amyloid ELISA A Reliable Way to Detect Endogenous Murine β-Amyloid. Teich, A.F, Patel, M., & Arancio, O. PLoS One (2013): 8(2):e55647

Differential Effects between γ–Secretase Inhibitors and Modulators on Cognitive Function in Amyloid Precursor Protein- Transgenic and Nontransgenic Mice. Mitani, Y., Yarimizu, J., Saita, K., Uchino, H., Akashiba, H., Shitaka, Y., Ni, K., & Matsuoka, N. J Neurosci (2012):32(6):2037-2050.

Temporal course of cerebrospinal fluid dynamics and amyloid accumulation in the aging rat brain from three to thirty months. Chiu, C., Miller, M.C., Caralopoulos, I.N., Worden, M.S., Brinker,T., Gordon, Z.N., Johanson, C.E., & Silverberg, G. D. Fluids Barriers CNS (2012): 9(1):3.

Reduction in BACE1 decreases body weight, protects against diet-induced obesity and enhances insulin sensitivity in mice. Meakin, P.J., Harper, A.J., Hamilton, D.L, Gallagher, J., McNeilly, A.D., Burgess, L.A., Vaanholt, L. M., Bannon, K.A., Latcham, J., Hussain, I., Speakman, J.R., Howlett, D.R., & Ashford, M. L. J. Biochem J (2012):441(1):285-296.

Amyloid-β42 Alters Apolipoprotein E Solubility in Brains of Mice with Five Familial AD Mutations. Youmans, K. L., Leung, S., Zhang, J., Maus, E., Baysac, K., Bu, G., Vassar, R., Yu, C., LaDu, M.J. J Neurosci Methods (2011): 196(1):51-59.

Rentinoic acid receptor-α signalling antagonises both intracellular and extracellular amyloid-β production and prevents neuronal cell death caused by amyloid-β. Jarvis, C. I., Goncalves, M.B., Clarke, E., Dogruel, M., Kalindijan, S.B., Thomas,

... Neurobiology ... Neurochemistry ... Neuropathology ... Neurobiology ... Neurochemistry ... Neuropathology ... Neurobiology ... Neurochemistry ... Neur

14 Bibliography

S.A., Maden, M., & Corcoran, J.P. (2010): 32(8):1246-1255. Amyloid-beta causes memory impairment by disturbing the JAK2/STAT3 axis in hippocampal neurons. Chiba, T., Yamada, M., Sasabe, K., Terashita, K., Shimoda, M., Matsuoka, M., Aiso, S. Mol Psychiatry (2009): 14(2): 206-222

Probes for senile plaque and neurofibrillary tangles Quinoline and Benzimidazole Derivatives: Candidate Probes for In Vivo Imaging of Tau Pathology in Alzheimer’s Disease. Okamura, N., Suemoto, T., Furumoto, S., Suzuki, M., Shimadzu, H., Akatsu, H., Yamamoto, T., Fujiwara, H., Nemoto, M., Maruyama, M., Arai, H., Yanai, K., Sawada, T. & Kudo, Y. J Neurosci (2005): 25(47):10857-10862. www.alphalabs.co.uk

Stryrylbenzoxazole Derivatives for In Vivo Imaging of Amyloid Plaques in the Brain. Okamura, N., Suemoto, T., Shimadzu, H., Suzuki, M., Shiomitsu, T., Akatsu, H., Yamamoto, T., Staufenbiel, M., Yanai, K., Arai, H., Sasaki, H., Kudo, Y., & Sawada, T. J Neurosci (2004):24(10):2535-2541.

Development of positron-emission tomography/single-photon emission computed tomography imaging probes for in vivo detection of beta-amyloid plaques in Alzheimer’s brains. Ono, M. Chem Pharm Bull (Tokyo). (2009):57(10):1029-1039.

Glutamate signalling Blockade of Gap Junction Hemichannel Suppresses Disease Progression in Mouse Models of Amyotrophic Lateral Sclerosis and Alzheimer’s Disease. Takeuchi, H., Mizoguchi, H., Doi, Y., Jin, S., Noda. M., Liang, J., Li, H., Zhou, Y., Mori, R., Yasuoka, S., Li, E., Parajuli, B., Kawanokuchi, J., Sonobe, Y., Sato. J., Yamanaka, K., Sobue, G., Mizuno, T., and Suzumnura, A. (2011): PLoS ONE 6(6): e21108

Development of a Highly Sensitive Cytotoxicity Assay System for CYP3A4-Mediated Metabolic Activation. Hosomi, H., Fukami, T., Iwamura, A., Nakajima, M., and Yokoi, T. Drug Metab Dispos .(2011): 39(8):1388-1395.

Phytochemicals A Microliter-Scale High-throughput Screening System with Quantum-Dot Nanoprobes for Amyloid-beta Aggregation Inhibitors. Ishigaki, Y., Tanaka, H., Akama, H., Ogara, T., Uwai, K., & Tokuraku, K. PLoS ONE (2013):8(8):e72992 doi:10.1371/journal.pone.0072992

Testosterone Deficiency Accelerates Neuronal and Vascular Aging of SAMP8 Mice: Protective Role of eNOS and SIRT1. Ota, H., Akishita, M., Akiyoshi, T., Kahyo, T., Setou, M., Ogawa, S., iijima, K., Eto, M., & Ouchi, Y. PLoS ONE (2012): 7(1):e29598. doi:10.1371/journal.pone.0029598

Resveratrol Ameliorates Muscular Pathology in the Dystrophic mdx Mouse, a Model for Duchenne Muscular Dystrophy. Hori, Y.S., Kuno, A., Hosoda, R., Tanno, M., Miura, T., Shimamoto, K. & Horio, Y. J Pharmacol. Exp. Ther (2011): 338(3):784-794

Effect of dietary resveratrol on the metabolic profile of nutrients in obese OLETF rats. Nagao, K., Jinnouchi, T., Kai, S. & Yanagita, T. Lipids Health Dis (2013):12(8) doi:10.1186/1476-511X-12-8

The dietary compound curcumin inhibits p300 histone acetyltransferase activity and prevents heart failure in rats. Morimoto. T., Sunagawa, Y., Kawamura, T., Takaya, T., Wada, H., Nagasawa, A., Komeda, M., Fujita, M., Shimatsu, A., Kita, T., & Hasegawa, K. J Clin Invest (2008):118(3):868-878

... Neurobiology ... Neurochemistry ... Neuropathology ... Neurobiology ... Neurochemistry ... Neuropathology ... Neurobiology ... Neurochemistry ... Neur

15 Bibliography

Phosphorylated alpha synuclein A quantitative study of α-synuclein pathology in fifteen cases of dementia associated with Parkinson disease. Armstrong, R.A., Kotzbauer, P.T., Perlmutter, J.S., Campbell, M.C., Hurth, K.M., Schmidt, R.E. & Cairns, N.J. J Neural Transm (2013) 10.1007/s00702-013-1084-z

α-Synuclein modulation of Ca2+ signalling in human neuroblastoma (SH-SY5Y) cell. Hettiarachchi, N.T., Parker, A., Research Dallas, M.L., Pennington, K., Hung, CC, Pearson, H.A., Boyle, J. P., Robinson, P., & Peers, C. J of Neurochem (2009): 111(5):1192-1201.

Accumulation of Phosphorylated α-Synuclein in Aging Human Brain. Saito, Y., Kawashima, A., Ruberu, N.N., Fujiwara, H., Koyama, S., Sawabe, M., Arai, T., Nagura, H., Yamanouchi, H., Hasegawa, M., Iwatsubo, T., & Murayama, S. J Neuropathol Exp Neurol (2003): 62(6):644-654

α-Synuclein is phosphorylated in synucleinpathy lesions. Fujiwara, H., Hasegawa, M., Dohmae, N., Kawashima, A., Masilah, E., Goldberg, M.S., Shen, J., Takio, K., & Iwatsubo, T. Nature Cell Biology (2002): 4(2):160-164

P2X4 IFN-gamma receptor signalling mediates spinal microglia activation driving neuropathic pain. Tsuda, M., Masuda, T., Kitano, J., Shimoyama, H., Tozaki-Saitoh, H., & Inoue, K. Proc. Natl Acad Sci (2009):106(19):8032-8037.

BDNF from microglia causes the shift in neuronal anion gradient underlying neuropathic pain. Coull. J. A., Begg, S., Boudreau, D., Boivin, D., Tsuda, M, Inoue, K, Gravel, C, Salter, M.W., & De Koninck, Y. Nature (2005):438(7070):1017-1021

P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury. Tsuda, M., Shigemoto-Mogami, Y., Koizumi, S., Mizokoshi, A., Kohsaka, S., Salter, M.W., & Inoue, K. Nature (2003): 424(6950):778-783.

Pikachurin Presynaptic Dystroglycan-Pikachurin Complex Regulates the Proper Synaptic Connection between Retinal Photoreceptor and Bipolar Cells. Omori, Y., Araki, F., Chaya, T., Kajimura, N., Irie, S., Terada, K., Muranishi, Y., Tsujii, T., Ueno, S., Koyasu, T, Tamaki, Y., Kondo, M., Amano, S., and Furukawa, T. J Neuroscience (2012): 32(18):6126-6137.

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