Hsp70 Antibody (OASE00332)

• Product Number: OASE00332
• Name: Hsp70 Antibody (OASE00332)
• Conjugation: Unconjugated
• NCBI Gene Id: 3303
• Host: Rabbit
• Clonality: Polyclonal
• 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 (1, 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 ATPbinding 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. Looking for more information on HSP70? Visit our new HSP70 Scientific Resource Guide at http://www.HSP70.com.
• Alias Symbols: HSP70 1, HSP70 2, HSP70.1, HSP72, HSP73, HSPA1, HSPA1A, HSPA1B
• Product Format: Liquid. PBS with 50% glycerol and 0.09% sodium azide.
• Reference: 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. Hung T.H., et al. (2001) Am J Pathol. 159: 1031-1043.
  8. Locke M. (2000) Cell Stress & Chaperones 5: 45-51.
  9. Ianaro A., et al. (2001) FEBS Lett. 508: 61-66.
  10.Trentin G.A. et al. (2001) J Biol Chem. 276: 13087-
  13095.
• Description of Target: 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, 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 ATPbinding 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.
• Reconstitution and Storage: Store at -20C. Shipping with Blue Ice or 4C.
• Specificity: Detects a ~70kDa. May cross-react with HSC70 at lower dilutions.
• Additional Information: Certificate of Analysis: A 1:1000 dilution of OASE00332 was sufficient for detection of HSP70 in 20 ug of HeLa cell lysate by ECL immunoblot analysis.
• Additional Information: Cellular Location: Cytoplasm.
• Application Info: WB: 1:1000
  IP: 1:100
  IHC: 1:100
  ICC/IF: 1:100
• Immunogen: Recombinant Full length HSP70 Protein
• Dilution: WB (1:1000), IHC (1:100), ICC/IF (1:100), IP (1:100); optimal dilutions for assays should be determined by the user.
• Storage Buffer: PBS pH7.4, 50% glycerol, 0.09% sodium azide
• Uniprot ID: P0DMV8/P0DMV9
• Protein Accession #: NP_005336.3
• Purification: Peptide Affinity Purified
• Gene Symbol: HSP70
• Predicted Species Reactivity: Human, Mouse, Rat, Beluga, Bovine, Dog, Fish, Carp, Guinea Pig, Hamster, Monkey, Pig, Sheep, Coral, Plant, Shark, L. amazonensis
• Application: WB, IHC, ICC, IF, IP, ELISA
• Image 1: Immunofluorescence
  
  Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Hsp70 Polyclonal Antibody. Tissue: Heat Shocked Cervical cancer cell line (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Hsp70 Polyclonal Antibody at 1:100 for 12 hours at 4C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Hsp70 Antibody. (C) Composite. Heat Shocked at 42C for 1h.
• Image 2: Immunofluorescence
  
  Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Hsp70 Polyclonal Antibody. Tissue: Heat Shocked Cervical cancer cell line (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Hsp70 Polyclonal Antibody at 1:100 for 12 hours at 4C. Secondary Antibody: APC Goat Anti-Rabbit (red) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Hsp70 Antibody. (C) Composite. Heat Shocked at 42C for 1h.
• Image 3: Immunohistochemistry
  
  Immunohistochemistry analysis using Rabbit Anti-HSP70 Polyclonal Antibody. Tissue: Inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Rabbit Anti-HSP70 Polyclonal Antibody at 1:1000 for 12 hours at 4C. Secondary Antibody: Biotin Goat Anti-Rabbit at 1:2000 for 1 hour at RT. Counterstain: Methyl Green at 200uL for 2 min at RT.
• Image 4: Immunohistochemistry
  
  Immunohistochemistry analysis using Rabbit Anti-HSP70 Polyclonal Antibody. Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Rabbit Anti-HSP70 Polyclonal Antibody at 1:50000 for 12 hours at 4C. Secondary Antibody: Biotin Goat Anti-Rabbit at 1:2000 for 1 hour at RT. Counterstain: Methyl Green at 200uL for 2 min at RT.
• Image 5: Western Blot
  
  Western blot analysis of Human, Rat brain cell lysates showing detection of HSP70 protein using Rabbit Anti-HSP70 Polyclonal Antibody. Load: 2 ug. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rabbit Anti-HSP70 Polyclonal Antibody at 1:10000 for 2 hours at RT. Secondary Antibody: Donkey Anti-Rabbit IgG: HRP for 1 hour at RT.
• Image 6: Western Blot
  
  Western blot analysis of Mouse Pam212 cells showing detection of HSP70 protein using Rabbit Anti-HSP70 Polyclonal Antibody. Load: 15 ugprotein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rabbit Anti-HSP70 Polyclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Donkey Anti-Rabbit IgG: HRP for 1 hour at RT.