Hsp70 Antibody (OASE00095)

• Product Number: OASE00095
• Name: Hsp70 Antibody (OASE00095)
• Clone: 2A4
• Isotype: IgM
• Conjugation: Unconjugated
• NCBI Gene Id: 3303
• Host: Mouse
• Clonality: Monoclonal
• Concentration: 1 mg/ml
• Gene Full Name: heat shock 70kDa protein 1A
• Description: 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.
• Alias Symbols: HSP70 1, HSP70 2, HSP70.1, HSP72, HSP73, HSPA1, HSPA1A, HSPA1B
• Reference: 1. Balashova N. et al. (2005) J Biol Chem 280:2186-96. 2. Boorstein W. R., Ziegelhoffer T. & Craig E. A. (1993) J. Mol. Evol.38 (1): 1-17.
  2. Rothman J. (1989) Cell 59: 591 -601.
  3. DeLuca-Flaherty et al. (1990) Cell 62: 875-887.
  4. Bork P., Sander C. & Valencia A. (1992) Proc. Nut1 Acad.
  Sci. USA 89: 7290-7294.
  5. Fink A.L. (1999) Physiol. Rev. 79: 425-449.
• Reconstitution and Storage: Store at -20C. Shipping with Blue Ice or 4C.
• Specificity: Detects ~70kDa. May detect HSP70, HSC70, and HSP72.
• Additional Information: Background Info: Detects several members of the heat shock protein 70kDa gene family including Hsp70, Hsc70, and following heat shock, Hsp72 from yeast, Drosophila, fish, mouse, avian, amphibian and human samples. IF staining of Hsp70 in heat shocked HeLa cells results in cytoplasmic staining.
• Additional Information: 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 (1). 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 (2). 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 (3). The structure of this ATP binding domain displays multiple features of nucleotide binding proteins (4).
  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 (5). 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.
• Immunogen: Human recombinant HSP70 overexpressed in E.coli
• Additional Information: Certificate of Analysis: 0.2 ug/mL was sufficient for detection of Hsp70 in 20ug of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgM:HRP as the secondary antibody.
• Dilution: WB (1:5000), IHC (1:100), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.
• Storage Buffer: PBS pH7.2, 50% glycerol, 0.09% sodium azide
• Uniprot ID: P0DMV8/P0DMV9
• Protein Accession #: NP_005336.3
• Purification: PEG Purified
• Gene Symbol: HSP70
• Predicted Species Reactivity: Human, Mouse, Rat, Amphibians, Chicken, Fish, Yeast, Drosophila
• Application: WB, IHC, ICC, IF, IP, AM
• Image 1: Immunofluorescence
  
  Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 2A4. Tissue: Cervical cancer cell line (HeLa). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody 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.
• Image 2: Western Blot
  
  Western Blot analysis of Rat Cell lysates showing detection of Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone 2A4. Load: 15 ug. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody at 1:5000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.