|Product Name||HSP70 Antibody|
Mouse Anti-Bovine HSP70 Monoclonal IgG1
|Species Reactivity||Human, Mouse, Rat, Bovine, Pig|
|Applications||, WB , ICC/IF , ELISA|
|Antibody Dilution||WB (1:500), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.|
|Conjugates||Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
AP (Alkaline Phosphatase)
HRP (Horseradish peroxidase)
|Storage Buffer||PBS pH7.4, 50% glycerol, 0.09% sodium azide|
|Shipping Temperature||Blue Ice or 4ºC|
|Purification||Protein G Purified|
|Specificity||Detects 70kDa. Does not cross react with HSC70.|
|Cite This Product||Mouse Anti-Bovine HSP70 Monoclonal, Clone 1.86 (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SMC-113)|
|Certificate of Analysis||1 µg/ml of SMC-113 was sufficient for detection of HSP70 in 20 µg of Hela lysate by colorimetric immunoblot analysis using goat anti-mouse IgG:HRP as the secondary antibody.|
|Alternative Names||HSP70 1 Antibody, HSP70 2 Antibody, HSP70.1 Antibody, HSP72 Antibody, HSPA1 Antibody, HSPA1A Antibody, HSPA1B Antibody|
|Research Areas||Cancer, Heat Shock|
|Scientific 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 50% 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. For more information visit our HSP70 Scientific Resource Guide at http://www.HSP70.com.
|References||1. Welch W.J. and Suhan J.P. (1986) J Cell Biol. 103: 2035-2050.
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.
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1.86 (SMC-113). Tissue: Cervical Cancer cell line (HeLa). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-113) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Nucleus, Cytoplasm. Magnification: 40X.
Western Blot analysis of Human HEK293 cell lysate showing detection of ~70 kDa Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone 1.86 (SMC-113). Lane 1: MW ladder. Lane 2: Human HEK293 lysate (20 µg). Load: 20 µg. Block: 5% milk + TBST for 1 hour at RT. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody (SMC-113) at 1:500 for 1 hour at RT. Secondary Antibody: HRP Goat Anti-Mouse at 1:100 for 1 hour at RT. Color Development: TMB solution for 5 min at RT. Predicted/Observed Size: ~70 kDa.
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