FluoProbes® FT-FP532A FluoProbes® 532A

Product Information

A great orange fluorophore for labeling biomolecules with fluorescent orange emission (alternative to A532)

Product name MW exc\em. max. mol. abs. Quantum cat.number (g·mol-1) (nm) (M-1cm-1) yield M+ (%) Fluoprobes® 532A Carboxyl group 765 646 (L) FP-BA6940, 1mg 532 / 553 115 000 90 Fluoprobes® 532A NHS 1081 743 FP-BA6950, 1mg (K) fl 90% Fluoprobes® 532A Maleimide 1063 768 (M) FP-BA6960, 1mg fl 3.8 ns Fluoprobes® 532A – Iodoacetamide CF260 = ε260/εmax 0.22

(V) FP-YE5260, 1mg CF280 = ε280/εmax 0.11 Fluoprobes® 532A – Azide

(V) FP-YE4980, 1mg Fluoprobes® 532A – Prot.Lab. Kit

(V) Inquire Other Fluoprobes® 550A products See related products M+: molecular mass of dye cation (MS)

Storage: (L): at +4°C (K) : at +4°C long term at –20°C) (M): at –20°C U F R 1,0 Fluoprobes® provide a full range of fluorophores to covers any applications, spanning from 390nm to ® 800nm. Fluoprobes dyes are designed for labeling biomolecules in advanced fluorescent detection 0,8 techniques. Applications include multiple labeling, FRET, Quenching, polarisation anisotropy FP®390 - Em. 0,6 FP®488 - Em. fluorescence, and life time resolved fluorescence, with protein as well as with nucleic acids, as well as FP®532 - Em. FP®547H - Em. FP®565 - Em. FP®594 - Em. dying materials. FP®647H - Em. 0,4 FP®682 - Em. Please see a presentation of selected most popular and remarkable FluoProbes labels in standard FP®782 - Em. applications (i.e. blue, green, orange, red, infrared), or at pages B51-B57 of the BioSciences catalogue, 0,2 and updated list of FluoProbes dyes NHS esters.

nm 0,0 350 450 550 650 750 850

Scientific and technical Information - Label

® Fluoprobes 532A label offers bright orange fluorescence (λexc./λem.: 532/553nm)

Absorption and emission spectra:  excited most efficiently in the range 515 - 545 nm i.e; by the 532 nm line of the frequency-doubled Nd:YAG laser.  very bright  high photostability  excellent water solubility*  carries a net electrical charge of –1 after coupling  compatible with standard filters for A532

*Both NHS and maleimide forms are soluble at 2.5mg/ml in water

Spectra of absorption (in water) + Contact your local distributor FluoProbes®, powered by [email protected]

P.1 FluoProbes® FT-FP532A Scientific and technical Information - Derivatives

Fluoprobes®532A is available as different derivatives, suiting standard chemistry methods (seek in the literature the right conjugation strategy for your application), and others (dying materials,...). It is supplied as a crystalline, solvent-free solid. Due to particular substituents FP532 is very hydrophilic. After coupling to a substrate the label carries a net electrical charge of -1.

Storage and General uses

Carboxylic derivatives are can be used for any kind of spectroscopy, and coupled to biomolecules by conventional chemistry, i.e. after activation at the carboxy group by EDC. Carboxylic derivatives are stored at ambient temperature and are stable for at least three years.

NHS-ester derivatives are suited for direct labeling of amino groups in proteins and aminated DNA/RNA. The chemical group N-hydroxysuccinimidyl (NHS) reacts specifically with primary (–NH2) and secondary amines (-NH-) (in fact on its deprotonated form) in aqueous phase or at pH 8 (compatible with pH7 to 10) in PBS buffer (other buffer devoid of amines are possible) at a ratio of 1-6 over amine content. I.e. amines present in proteins (Lys aminoacid) and in a lower proportion on NH2 located in terminal peptidic chains. The reaction competes with hydrolysis that increases with pH, and with the high dilutions of the molecule that should be labeled. Please refer to the literature, or the technical sheet FT-BA680 (NHS-FluoProbes labels) for standard protocols. NHS-esters can be stored at 0-4°C, stable for several months, or at –20°C for long term. They should be protected from moisture and light.

Maleimide derivatives are suited for labeling of thiol groups of proteins or other molecules, e.g. specific labeling of cysteine. The maleimide group reacts very specifically with sulfhydryls –SH at neutral pH 6.5-7. The reaction is rapid (a few minutes for cysteine), but in the absence of –SH, maleimide stay well stable. In usual conditions, one should start with a ratio of 10-20 moles of maleimide per mole of protein. Please refer to the literature, or the technical sheet FT-BA681 (Maleimide-FluoProbes labels) for standard protocols. Maleimide derivatives can be stored at 0-4°C, stable for several months, or at –20°C for long term. They should be protected from moisture and light.

Hydrazide derivatives are available on inquire

You also may ask Protein labeling kits, already prepared Fluoprobes conjugates (see related products), and custom labeling. Standard protocols (*) Protocol 1 : antibody labeling with NHS ester See the technical sheet FT-BA6800. This simple and quick standard protocol labels polyclonal and monoclonal purified antibodies for immunodetection applications. It suits also most proteins and peptides (*).

Protocol 2 : Incorporation of aa-dUTP by Reverse Transcription See the technical sheet FT-BA68000. AminoAllyl-UTP (aa-UTP) is incorporated in nucleic acids using a DNA polymerase (PCR, Nick translation) for subsequent labeling by NHS-FluoProbes®dye.

Protocol 3 – protein labeling with maleimide See the technical sheet FT-BA68100. Fluoprobes® dye maleimide is suited for labeling of proteins at cysteine sites.

(*)A calibration of dye/biomolecule ratio may be needed to optimize the labeling level depending on molecule and application, i.e. adjust concentration weight of the FluoProbes® dye / weight of protein or peptide. Then the parameters of the detection instrument should also be sat properly for FluoProbes dye (see above/label).

Other FluoProbes532A labeled products:

Fluoprobes®532A – Streptavidin FP-CA5590, 1mg (L) Biotin – FP532A FP-BA6970 Custom labeling Inquire Phalloidin – FP532A FP-YE5190 Nucleotides – Fluoprobes532A Inquire: examples: N6-(6-Amino)Hexyl-ATP-FP532AFP-FQ5030 8-((6-Amino)Hexyl)-Amino-ATP-FP532A FP-FQ4720 EDA-ATP – FP532A FQ-FQ5750 EDA-ADP – FP532A FP-FQ5660 EDA-APPNHP (EDA-AMPPNP) FP-FQ7280 Aminoallyl-UTP - FP532A FP-FQ5550 Aminoallyl-UTP – FP532A FP-IL7550 dUTP – FP532A - dCPT – FP532A FP-FQ5310 Complete list of FluoProbes dyes NHS esters at http://www.interchim.fr/ft/F/FPlistN.pdf .

References for FluoProbes532A labeled products: Guillaume Clavé et al; Org. Biomol. Chem., 2008, DOI: 10.1039/b807263a; "A novel heterotrifunctional peptide-based cross-linking reagent for facile access to bioconjugates. Applications to peptide fluorescent labelling and immobilisation"; Abstract; Article [preparation of fluorescent tripod-functionalised surfaces which may be useful in developing new peptide microarrays and related immunosensors]

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P.2 FluoProbes® FT-FP532A Ordering information

Catalog size quantities and prices may be found at http://www.interchim.com Please inquire for higher quantities (availability, shipment conditions).

For any information, please ask : FluoProbes® / Interchim; Hotline : +33(0)4 70 03 73 06

Disclaimer : Materials from FluoProbes® are sold for research use only, and are not intended for food, drug, household, or cosmetic use. FluoProbes® is not liable for any damage resulting from handling or contact with this product. Rev. H04E/O

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