Monoclonal Antibody to HSPA8 / HSC70

AM03141PU-N OriGene Technologies Inc. OriGene EU

Acris Antibodies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-888-267-4436 Phone: +49-5221-34606-0 Fax: +1-301-340-8606 Fax: +49-5221-34606-11 [email protected] [email protected]

Monoclonal Antibody to HSPA8 / HSC70 - Aff - Purified Alternate names: HSP73, HSPA10, Heat shock 70 kDa protein 8, Heat shock cognate 71 kDa protein Catalog No.: AM03141PU-N Quantity: 50 µg Concentration: 1.0 mg/ml Background: Hsp70 genes encode abundant heat-inducible 70-kDa hsps (hsp70s). In most eukaryotes hsp70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 5O% identity (2). The N-terminal two thirds of hsp70s are more conserved than the C-terminal third. Hsp70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides (3). When hsc70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44 kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half (4). The structure of this ATP binding domain displays multiple features of nucleotide binding proteins (5). All hsp70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the hsp70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein (6). The universal ability of hsp70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport. Uniprot ID: P11142 NCBI: NP_006588 GeneID: 3312 Host / Isotype: Mouse / IgG2a Clone: BB70 Immunogen: Chicken Hsp70/Hsp90 complex (1) Format: State: Liquid purified IgG fraction Purification: Affinity Chromatography on Protein G Buffer System: PBS pH 7.2 Preservatives: 0.09% Sodium Azide Stabilizers: 50% Glycerol

For research and in vitro use only. Not for diagnostic or therapeutic work. OG/20160219 Material Safety Datasheets are available at www.acris-antibodies.com or on request. Acris Antibodies is now part of the OriGene family. Learn more at www.origene.com 1 / 3 AM03141PU-N: Monoclonal Antibody to HSPA8 / HSC70 - Aff - Purified Applications: Western blot (7): 1 µg/ml was sufficient for detection of hsp70/Hsc70 in 20 µg of Heat Schocked HeLa lysate by colorimetric Immunoblot analysis using Goat anti-Mouse IgG -HRP as the secondary antibody. Immunoprecipitation (8). Immunohistochemistry (9). Immunocytochemistry/Immunofluorescence: This antibody clone BB70 is reported to stain Hsp70 + Hsc70 in nuclear smears of Rat hepatocytes by Immunocytochemistry (10). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user. Specificity: Detects 72 and 73kDa proteins corresponding to the Molecular Mass of inducible HSP and HSC70 on SDS PAGE Immunoblots. Species: Human, mouse, rat, sheep, dog, beluga, bovine, fish (carp, rainbow trout, Chinook/Chum/Coho salmon), pacific oyster, guinea pig, scallop pig, hamster, rabbit, chicken, Artemia, Xenopus, Drosophila, and yeast. Other species not tested. Storage: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: one year from despatch. Product Citations: Purchased from Acris: 1. Vavilis T, Delivanoglou N, Aggelidou E, Stamoula E, Mellidis K, Kaidoglou A, et al. Oxygen- Glucose Deprivation (OGD) Modulates the Unfolded Protein Response (UPR) and Inflicts Autophagy in a PC12 Hypoxia Cell Line Model. Cell Mol Neurobiol. 2015 Aug 4. PubMed PMID: 26239244. 2. Stamoula E, Vavilis T, Aggelidou E, Kaidoglou A, Cheva A, Mellidis K, et al. Low Dose Administration of Glutamate Triggers a Non-Apoptotic, Autophagic Response in PC12 Cells. Cell Physiol Biochem. 2015;37(5):1750-8. doi: 10.1159/000430250. Epub 2015 Nov 13. PubMed PMID: 26584276. Originator or purchased from resellers: 1. Kang Y, Taldone T, Patel HJ, Patel PD, Rodina A, Gozman A, et al. Heat shock protein 70 inhibitors. 1. 2,5'-thiodipyrimidine and 5-(phenylthio)pyrimidine acrylamides as irreversible binders to an allosteric site on heat shock protein 70. J Med Chem. 2014 Feb 27;57(4):1188-207. doi: 10.1021/jm401551n. Epub 2014 Feb 18. PubMed PMID: 24548207. 2. Taldone T, Kang Y, Patel HJ, Patel MR, Patel PD, Rodina A, et al. Heat shock protein 70 inhibitors. 2. 2,5'-thiodipyrimidines, 5-(phenylthio)pyrimidines, 2-(pyridin-3-ylthio)pyrimidines, and 3-(phenylthio)pyridines as reversible binders to an allosteric site on heat shock protein 70. J Med Chem. 2014 Feb 27;57(4):1208-24. doi: 10.1021/jm401552y. Epub 2014 Feb 18. PubMed PMID: 24548239. 3. Rodina A, Taldone T, Kang Y, Patel PD, Koren J, Yan P, et al. Affinity purification probes of potential use to investigate the endogenous Hsp70 interactome in cancer. ACS Chem Biol. 2014 Aug 15;9(8):1698-705. doi: 10.1021/cb500256u. Epub 2014 Jun 17. PubMed PMID: 24934503. 4. Rodina A, Patel PD, Kang Y, Patel Y, Baaklini I, Wong MJ, et al. Identification of an allosteric pocket on human hsp70 reveals a mode of inhibition of this therapeutically important protein. Chem Biol. 2013 Dec 19;20(12):1469-80. doi: 10.1016/j.chembiol.2013.10.008. Epub 2013 Nov 14. PubMed PMID: 24239008. General Readings: 1. Zho J. (1998) Cell 94 : 471-480 2. Boorstein W. R., Ziegelhoffer T. & Craig E. A. (1993) J. Mol. Evol.38 (1): 1-17. 3. Rothman J. (1989) Cell 59: 591 -601. 4. DeLuca-Flaherty et al. (1990) Cell 62: 875-887. 5. Bork P., Sander C. & Valencia A. (1992) Proc. Nut1 Acad. Sci. USA 89: 7290-7294. 6. Fink A.L. (1999) Physiol. Rev. 79: 425-449.

Human colon tissue stained (brown) in inflammatory cells.? Shown at a 1:100,000 dilution.

Hsp70/Hsc70 detection in paraffin embedded liver sections and nuclear sections of rat hepatocytes by staining with BB70.? First pictures in series show two hours after exposure to stress, the second shows the control.