Heat Shock Protein 70, Recombinant Human (Hsp70)

Hsp70 genes encode abundant heat-inducible 70kD 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 44kDa 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 \nintermediates 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 \nprotein transport.\n\nSource: \nProtein with ATPase activity, his-tagged, Recombinant Hsp70 cloned from a human cDNA library and expressed in E.coli.\n\nApplications:\nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

H1830-95T7

Size

50ug

Applications

WB

Form

Supplied as a liquid in sodium phosphate, pH 7.2, (20mM), 150mM sodium chloride, 10% glycerol, 200mM Imidazole.

Purity

~90% (SDS-PAGE).

References

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. \n6. Fink, A.L. (1999) Physiol. Rev. 79: 425-449. 7. Smith, D.F., et al., (1993) Mol. Cell. Biol. 13(2):869-876. 8. Prapapanich, V., et al., (1996) Mol. Cell. Biol. 16(11):6200-6207. 9.Fernandez-Funez et al., (2000) Nature 408(6808):101-106.