Mouse Hsp70 Monoclonal Antibody | anti-HSPA1A antibody

Hsp70 Antibody

Cat. Num. AAA801280

• Gene Names: HSPA1A; HSP72; HSPA1; HSP70I; HSP70-1; HSP70-1A; HEL-S-103
• Reactivity: Human, Mouse, Rat, Bovine, C.elegans, Canine, Chicken, Drosophilia, Carp, Guinea pig, Hamster, Monkey, Pig, Rabbit, Sheep
• Applications: Western Blot, Immunoprecipitation, ELISA, Immunocytochemistry, Immunohistochemistry, Flow Cytometry, Functional Assay
• Synonyms: Hsp70; Monoclonal Antibody; Hsp70 Antibody; Hsp70 1; Hsp70 2; Hsp70.1; Hsp72; HSPA1; HSPA1A; HSPA1B; Hsp 70; anti-HSPA1A antibody

• Host: Mouse
• Reactivity: Human, Mouse, Rat, Bovine, C.elegans, Canine, Chicken, Drosophilia, Carp, Guinea pig, Hamster, Monkey, Pig, Rabbit, Sheep
• Clonality: Monoclonal
• Isotype: IgG1
• Clone Number: C92F3A-5
• Form/Format: Protein G Purified
• Concentration: 1mg/mL (varies by lot)
• Sequence Length: 641

• Applicable Applications for anti-HSPA1A antibody: Western Blot (WB), Immunoprecipitation (IP), ELISA (EIA), Immunocytochemistry (ICC), Immunohistochemistry (IHC), Flow Cytometry (FC/FACS), IEM
• Application Notes: 1 ug/ml was sufficient for detection of Hsp70 in 20ug of Hela cell lysate
• Certificate of Analysis: 1 ug/mL was sufficient for detection of Hsp70 in 20ug of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.
• Storage Buffer: PBS pH7.2, 50% glycerol, 0.09% sodium azide.
• Preparation and Storage: -20 degree C

Immunohistochemistry (IHC)

(Immunohistochemistry analysis using Mouse Anti-Hsp70 Monoclonal Antibody, Clone C92. Tissue: colon carcinoma. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody at 1:10000 for 12 hours at 4 degree C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 ul for 2 minutes at RT. Localization: Inflammatory cells. Magnification: 40x.)

Immunocytochemistry/Immunofluorescence (ICC/IF)

(Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Hsp70 Monoclonal Antibody, Clone C92. Tissue: Heat Shocked Melanoma cells. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody at 1:1000 for 16 hours at RT. Secondary Antibody: Biotin Goat Anti-Mouse. Courtesy of: Dr. Ewa Malusecka, Maria Sklodowska-Curie Memorial Cancer Centre and Inst. Of Oncology, Poland.)

Immunohistochemistry (IHC)

(Immunohistochemistry analysis using Mouse Anti-Hsp70 Monoclonal Antibody, Clone C92. Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody at 1:10000 for 12 hours at 4 degree C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 ul for 2 minutes at RT. Localization: Inflammatory cells. Magnification: 40x.)

Western Blot (WB)

(Western Blot analysis of Human cell lysates from various cell lines showing detection of Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone C92. Load: 15 ug. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.)

Immunoflurescence (IF)

(Fluorescence Activated Cell Sorting analysis using Mouse Anti-Hsp70: FITC Monoclonal Antibody, Clone C92. Tissue: Heat Shocked CD3+ CD8+ T cells . Species: Mouse. Primary Antibody: Mouse Anti-Hsp70: FITC Monoclonal Antibody at 1:1000. Courtesy of: Cheryl Cameron, Vaccine and Gene Therapy Instit. Florida.)

Related Product Information for anti-HSPA1A antibody

Background Info: Detects a ~70kDa protein corresponding to the molecular mass of inducible Hsp70 on SDS PAGE immunoblots. The mapped epitope is in the region of amino acid residues 436-503. Does not cross-react with Hsc70 (Hsp73).

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 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.

Product Categories/Family for anti-HSPA1A antibody

Chaperones; Heat Shock; Trafficking

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. 7. Galan A., et al. (2000) J. Biol. Chem. 275: 11418-11424. 8. Kondo T., et al. (2000) J. Biol. Chem. 275: 8872-8879. 9. Misaki T., et al. (1994) Clin. Exp. Immun. 98: 234-239. 10. Pockley A.G., et al. (1998) Immunol. Invest. 27: 367-377. 11. Moon I.S., et al. (2001) Cereb Cortex 11(3): 238-248. 12. Dressel et al. (2000) J. Immunol. 164: 2362-2371. 13. Verma A.K., et al. (2007) Fish and Shellfish Immunology. 22(5): 547-555. 14. Banduseela V.C., et al. (2009) Physiol Genomics. 39(3): 141-159.
1. Xu, Y. et al. (2009). 2,3-Dihydrowithaferin A-3β-O-sulfate, a new potential prodrug of withaferin A from aeroponically grown Withania somnifera. Bioorganic & Medicinal Chemistry. 17 (6), 2210-2214. doi:10.1016/j.bmc.2008.10.091
2. Fernndez-Llama, P. et al. (2010). Tamm-Horsfall protein and urinary exosome isolation. Kidney International. 77, 736-742. doi:10.1038/ki.2009.550
3. Marubayashi, S. et al. (2010). HSP90 is a therapeutic target in JAK2-dependent myeloproliferative neoplasms in mice and humans. J Clin Invest. 120 (10), 3578-3593. doi:10.1172/JCI42442
4. Olkku, A., Leskinen, J.J., Lammi, M.J., Hynynen, K., and Mahonen, A. (2010). Ultrasound-induced activation of Wnt signaling in human MG-63 osteoblastic cells. Bone. 47 (2), 320-330.
5. Aare, S. et al. (2011). Mechanisms underlying the sparing of masticatory versus limb muscle function in an experimental critical illness model. Physiol Genomics. 43 (24), 1334-1350. doi: 10.1152/physiolgenomics.00116.2011.

NCBI and Uniprot Product Information

• NCBI GI #: 194248072
• NCBI GeneID: 3303
• NCBI Accession #: NP_005336.3
• NCBI GenBank Nucleotide #: NM_005345.5
• UniProt Accession #: P08107; P19790; Q5JQI4; Q5SP17; Q9UQL9; Q9UQM0; B4E3B6
• Molecular Weight: 63,937 Da
• NCBI Official Full Name: heat shock 70 kDa protein 1A/1B
• NCBI Official Synonym Full Names: heat shock 70kDa protein 1A
• NCBI Official Symbol: HSPA1A
• NCBI Official Synonym Symbols: HSP72; HSPA1; HSP70I; HSP70-1; HSP70-1A; HEL-S-103
• NCBI Protein Information: heat shock 70 kDa protein 1A/1B; HSP70-1/HSP70-2; heat shock 70kD protein 1A; heat shock-induced protein; heat shock 70 kDa protein 1/2; epididymis secretory protein Li 103; dnaK-type molecular chaperone HSP70-1
• UniProt Protein Name: Heat shock 70 kDa protein 1A/1B
• Protein Family: HSP70 co-chaperone
• UniProt Gene Name: HSPA1A
• UniProt Synonym Gene Names: HSPA1; HSX70; HSPA1B; HSP70-1/HSP70-2; HSP70.1/HSP70.2
• UniProt Entry Name: HSP71_HUMAN

NCBI Description

This intronless gene encodes a 70kDa heat shock protein which is a member of the heat shock protein 70 family. In conjuction with other heat shock proteins, this protein stabilizes existing proteins against aggregation and mediates the folding of newly translated proteins in the cytosol and in organelles. It is also involved in the ubiquitin-proteasome pathway through interaction with the AU-rich element RNA-binding protein 1. The gene is located in the major histocompatibility complex class III region, in a cluster with two closely related genes which encode similar proteins. [provided by RefSeq, Jul 2008]

Uniprot Description

HSP70: a critical chaperone protein that has a high affinity for unfolded polypeptide chains. It binds extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In cooperation with other chaperones, hsp70 stabilizes preexistent proteins against aggregation and mediates the folding of newly translated polypeptides in the cytosol as well as within organelles. Mitochondrial HSP70 is crucial to the import process: mutant forms of HSP70 fail to import precursor proteins. Is anti-apoptotic in sympathetic neurones and mediates this effect primarily by suppressing c-Jun transcriptional signalling. Interacts with tau protein and mediates proper folding of tau. Can promote the degradation of tau protein. Triptolide, a potential therapeutic agent for progression/metastasis of pancreatic cancer, causes pancreatic cancer cell death by induction of apoptosis, an effect mediated by the inhibition of HSP70.

Protein type: Chaperone; Motility/polarity/chemotaxis; Heat shock protein

Chromosomal Location of Human Ortholog: 6p21.3

Cellular Component: signalosome; focal adhesion; mitochondrion; endoplasmic reticulum; inclusion body; ribonucleoprotein complex; cytosol; nucleoplasm; centriole; perinuclear region of cytoplasm; cytoplasm; nuclear speck; nucleus; vesicle; ubiquitin ligase complex

Molecular Function: viral receptor activity; protein binding; enzyme binding; G-protein-coupled receptor binding; heat shock protein binding; ubiquitin protein ligase binding; double-stranded RNA binding; unfolded protein binding; ATPase activity; protein N-terminus binding; ATPase activity, coupled; ATP binding

Biological Process: negative regulation of cell proliferation; entry of virus into host cell; protein stabilization; positive regulation of erythrocyte differentiation; mRNA catabolic process; protein refolding; negative regulation of protein ubiquitination; gene expression; negative regulation of cell growth; response to unfolded protein; negative regulation of apoptosis