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Heat Shock Protein 70 (HSP70) BioAssay(TM) ELISA Kit

Cat no: H1834-37

Supplier: United States Biological
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Heat Shock Protein 70 (Hsp70) BioAssay(TM) ELISA Kit is a colorimetric sandwich immunoassay for the quantitative determination of inducible Heat Shock Protein (Hsp70) in serum and plasma samples of human, mouse and rat origin. It does not detect other Hsp70 family members such as Hsc70 (Hsp73), rp78, DnaK (E. coli), or Hsp71 (M. tuberculosis). The Hsp70 family of heat shock proteins contains multiple homologs ranging in size from 66-78 kDa, and are the eukaryotic equivalents of the bacterial DnaK. The most studied Hsp70 members include the cytosolic stress-induced Hsp70 (Hsp72), the constitutive cytosolic Hsc70 (Hsp73), and the ER-localized BiP (Grp78). Hsp70 family members contain highly conserved N-terminal ATP-ase and C-terminal protein binding domains. Binding of peptide to Hsp70 is assisted by Hsp40, and stimulates the inherent ATPase activity of Hsp70, facilitating ATP hydrolysis and enhanced peptide binding. Hsp70 nucleotide exchange and substrate binding coordinates the folding of newly synthesized proteins, the re-folding of misfolded or denatured proteins, coordinates trafficking of proteins across cellular membranes, inhibits protein aggregation, and targets the degradation of proteins via the proteasomal pathway. Hsp70 is a molecular chaperone whose expression is induced upon exposure of the cell or organism to conditions of stress. It prevents protein aggregation and promotes the refolding of proteins that become damaged in response to environmental insults, pathogens, and disease. Its activity is essential for cellular survival and recovery under stress conditions, as well as for the maintenance of normal cellular function under non-stress conditions (1-3). Hsp70 has been implicated to play a role in a variety of disease and physiological processes such as hyperthermia (4), hypertension (5), toxic exposure to chemical agents (6), hypoxia (7), ischemia (8,9), inflammation (10), autoimmunity (5, 11), apoptosis (12, 13), cancer (13), organ transplantation (14) and bacterial (15,16) and viral (17) infections. Hsp70 is also a key regulator of many normal physiological processes including aging (12,18), spermatogenesis (19,20), menstruation (21), and physical activity such as exercise (22). This kit is designed to evaluate and monitor Hsp70 in these processes, providing a key research tool to understand the role of Hsp70 in physiology and disease. \ Principle of the Assay: Samples and standards are added to wells coated with a monoclonal antibody specific for Hsp70. After incubation, the plate is washed, leaving only bound Hsp70 on the plate. A yellow solution of polyclonal antibody specific for Hsp70 is then added. This binds the Hsp70 captured on the plate. After incubation, the plate is washed to remove excess antibody. A blue solution of HRP conjugate is added to each well, binding to the antibody. The plate is again incubated. The plate is washed to remove excess HRP conjugate. TMB substrate solution is added. and the HRP-catalyzed reaction generates blue color in the solution. Stop solution is added to stop the substrate reaction. The resulting yellow color is read at 450nm. The amount of signal is directly proportional to the level of Hsp70 in the sample. Sensitivity: 0.09ng/ml Range: 0.20-12.5ng/ml Kit Components: H1834-37A: Microtiter Plate 1x96 wells, 12 x 8 removable strips. Pre-coated clear wells coated with Hsp70 monoclonal antibody. H1834-37B: Hsp70 Standard, 1x25ul. One vial containing 10ug/ml of recombinant human Hsp70. H1834-37C: Assay Buffer, 1x50ml. Phosphate buffered saline containing BSA and detergent. H1834-37D: Wash Buffer 20X, 1x100ml. Tris buffered saline containing detergents. H1834-37E: Hsp70 Pab, 1x10ml. A yellow solution of rabbit polyclonal antibody specific for Hsp70. H1834-37F: Pab (HRP), 1x10ml. A blue solution of goat anti-rabbit IgG conjugated to horseradish peroxidase. H1834-37G: TMB Substrate, 1x10ml. A solution of 3,3
Catalogue number: H1834-37
Reactivities: Human, Mouse, Rat
Applications: ELISA
Size: 1Kit
References: 1. Mayer, M.P., et al., Biol. Chem. 379: 261-268 (1998). 2. Bukau, B., et al., Cell 125: 443-451 (2006). 3. Mayer, M.P. & Bukau, B., Cell Mol. Life Sci. 62: 670-684 (2005). 4. Li, G.C., et al., Int. J. Hyperthermia 11: 459-488 (1995). 5. Lovis, C., et al., Ren. Fail. 16: 179-192 (1994). 6. Witzmann, F.A., et al., Electrophoresis 17: 198-202 (1996). 7. Chen, H.C., et al., Kidney Int. 56: 1270-1273 (1999). 8. Gray, C.C., et al., Int. J. Biochem. Cell. Biol. 31: 559-573 (1999). 9. Perdrizet, G.A., et al., Ann NY Acad Sci. 874: 320-325 (1999). 10. Jacquier-Sarlin, M.R., et al., Experientia 50: 1031-1038 (1994). 11. Feige, U., et al., EXS 77: 359-373 (1996). 12. Gabai, V.L., et al., FEBS Lett 438: 1-4 (1998). 13. Jaattela, M., Exp. Cell Res. 248: 30-43 (1999). 14. Mehta, N.K., et al., J. Surg. Res. 70: 151-155 (1997). 15. Delogu, G., et al., J. Crit. Care 12: 188-192 (1997). 16. Nishimura, H., et al., Cell Stress Chap. 2: 50-59 (1997). 17. Santoro, M.G., Experientia 50: 1039-1047 (1994). 18. Heydari, A.R., et al., Experientia 50: 1092-1098 (1994). 19. Eddy, E.M., Rev. Reprod. 4: 23-30 (1999). 20. Dix, D.J. & Hong, R.L., Adv. Exp. Med. Biol. 444: 137-143 (1998). 21. Komatsu, T., et al., J. Clin. Endocrinol. Metab. 82: 1385-1389 (1997). 22. Kilgore, J.L., et al., Can. J. Appl. Physiol. 23: 245-260 (1998).

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