HSP70/HSPA1A [ac Lys77] Antibody - BSA Free

Western Blot: HSP70/HSPA1A [ac Lys77] Antibody [NBP2-61122] - Western blot analysis of Human Cervical cancer cell line (HeLa) lysate showing detection of ~70 kDa HSP70/HSPA1A protein using Rabbit Anti-HSP70/HSPA1A Polyclonal Antibody (NBP2-61122). Lane 1: Molecular Weight Ladder (MW). Lane 2: Cervical Cancer cell line (HeLa) lysate. Lane 3: H2O2 Cervical Cancer cell line (HeLa) lysate. Load: 10 ug. Block: 5% Skim Milk in 1X TBST. Primary Antibody: Rabbit Anti-HSP70/HSPA1A Polyclonal Antibody (NBP2-61122) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Rabbit HRP:IgG at 1:3000 for 1 hour at RT. Color Development: ECL solution for 5 min at RT. Predicted/Observed Size: ~70 kDa.

Immunocytochemistry/Immunofluorescence: HSP70/HSPA1A [ac Lys77] Antibody [NBP2-61122] - analysis using Rabbit Anti-HSP70 Acetyl Lys77 Polyclonal Antibody. Tissue: Embryonic kidney cells (HEK293) cultured overnight with 50 uM H2O2. Species: Human. Fixation: 5% Formaldehyde for 5 min. Primary Antibody: Rabbit Anti-HSP70 Acetyl Lys77 Polyclonal Antibody at 1:60 for 30-60 min at RT. Secondary Antibody: Goat Anti-Mouse Alexa Fluor 488 at 1:1500 for 30-60 min at RT. Counterstain: Phalloidin Alexa Fluor 633 F-Actin stain; DAPI (blue) nuclear stain at 1:250, 1:50000 for 30-60 min at RT. Localization: Cytoplasmic. Magnification: 20X (2X Zoom). (A) DAPI (blue) nuclear stain. (B) Phalloidin Alex Fluor 633 F-Actin stain. (C) HSP70 Antibody (Acetyl Lys77). (D) Composite. Courtesy of: Dr. Robert Burke, University of Victoria.

Immunohistochemistry-Paraffin: HSP70/HSPA1A [ac Lys77] Antibody [NBP2-61122] - Tissue: Colon Cancer. Species: Human. Primary Antibody: Rabbit Anti-HSP70 (Acetyl Lys77) Polyclonal Antibody (NBP2-61122) at 1:50 for 30 min at RT. Counterstain: Hematoxylin. Magnification: 20X. HRP-DAB Detection.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ELISA, ICC/IF, IHC, IHC-Fr (-)
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 0.5 mg/ml

HSP70/HSPA1A [ac Lys77] Antibody - BSA Free Summary

• Description: Novus Biologicals Rabbit HSP70/HSPA1A [ac Lys77] Antibody - BSA Free (NBP2-61122) is a polyclonal antibody validated for use in IHC, WB, ELISA and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Synthetic peptide from the N-terminal of Human HSP70 Acetyl Lys77 (1-100 aa), conjugated to Keyhole Limpet Haemocyanin (KLH).
• Modification: ac Lys77
• Localization: Cytoplasm, Nucleus
• Specificity: Binds to acetylated lysine 77 on HSP70. Detects 70 kDa band.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HSPA1A
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:60
  • Immunohistochemistry 1:50
  • Immunohistochemistry-Paraffin
  • Western Blot 1:1000
• Application Notes: A 1:1000 dilution of this antibody was sufficient for detection of HSP70 Acetyl Lys77 in 10 ug of human HeLa cell lysates by ECL immunoblot analysis using goat anti-rabbit IgG:HRP as the secondary antibody.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.4), 50% Glycerol
• Preservative: 0.09% Sodium Azide
• Concentration: 0.5 mg/ml
• Purity: Peptide affinity purified

Alternate Names for HSP70/HSPA1A [ac Lys77] Antibody - BSA Free

• dnaK-type molecular chaperone HSP70-1
• FLJ54303
• FLJ54370
• FLJ54392
• FLJ54408
• FLJ75127
• Heat shock 70 kDa protein 1/2
• heat shock 70 kDa protein 1A/1B
• heat shock 70kD protein 1A
• heat shock 70kDa protein 1A
• heat shock-induced protein
• HSP70
• HSP70.1/HSP70.2
• HSP70-1
• HSP70-1/HSP70-2
• HSP70-1A
• HSP70I
• HSP72
• HSPA1
• HSPA1A
• HSPA1B

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: HSPA1A