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Technical Data Sheet

HM4381

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Hsp60 (N-terminal region)
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms
60 kDa

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Immunocytochemical labeling of Hsp60 in mitochondria in paraformaldehyde-fixed and NP40-permeabilized A7r5 cells. The cells were labeled with mouse monoclonal Hsp60 (HM4381). The antibody was detected using goat anti-mouse DyLight® 488.

Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Heat shock proteins (Hsp) are a family of highly conserved proteins that include both constitutively expressed (Hsp60, Hsp70, and Hsp90) and stress-induced (Hsp27 and Hsp72) proteins. Hsp60 is a mitochondrial protein that promotes protein folding and facilitates proteolytic degradation of misfolded or denatured proteins in the mitochondria. Hsp10 interacts with Hsp60 to regulate its substrate binding and ATPase activity. In HeLa and Jurkat mitochondria, Hsp60 associates with caspase-3 to form a complex that dissociates and releases from the mitochondria during apoptosis. Hsp60 accelerates the maturation of procaspase-3 through its ATP-dependent “foldase” activity. In addition to its protein folding activity, Hsp60 can bind the toll-like receptor-4 complex leading to production of TNFα and stimulation of a pro-inflammatory response in macrophages. Thus, the protein folding function of Hsp60 is involved in protein folding in both normal and apoptotic cells, while release of Hsp60 during necrosis is thought to stimulate a pro-inflammatory response.


Background References
Venner, T.J. et al. (1990) DNA Cell Biol. 9(8):545.
Cappello, F. et al. (2008) Cancer Biol Ther. 7(6):801.
Grundtman, C. et al. (2011) Arterioscler Thromb Vasc Biol. 31(5):960.
Immunogen
Clone M438 was generated from a recombinant protein corresponding to amino acid residues in the N-terminal region of human Hsp60. This sequence has high homology to similar regions in rat and mouse Hsp60.
Buffer and Storage
Mouse monoclonal purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody detects a 60 kDa* protein corresponding to the apparent molecular mass of Hsp60 on SDS-PAGE immunoblots of human Jurkat and rat A7r5 cells. In immunocytochemistry, anti-Hsp60 specifically stains mitochondria in paraformaldehyde fixed and NP-40 permeabilized cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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