HSP70/HSPA1A Antibody (3A3)

Western Blot: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Analysis of 20ul of gill tissue lysates from the salt marsh mussel, Guekensia demissa, isolated from various coves in Rhode Island (indicated above the lanes).

Immunocytochemistry/Immunofluorescence: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Analysis of Heat Shock Protein 70 (Hsp70) (green) in HeLa and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a Hsp70 monoclonal antibody, at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.

Immunohistochemistry-Paraffin: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Both normal and cancer biopsies of deparaffinized human Breast carcinoma tissue.

Western Blot: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a Hsp70 monoclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:20,000 for at least 1 hour.

Immunocytochemistry/Immunofluorescence: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Analysis of Heat Shock Protein 70 using Heat Shock Protein 70 Monoclonal antibody (3A3) shows staining in NIH-3T3 cells. Heat Shock Protein 70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown.

Immunocytochemistry/Immunofluorescence: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Analysis of Heat Shock Protein 70 using Heat Shock Protein 70 Monoclonal antibody (3A3) shows staining in MCF-7 cells. Heat Shock Protein 70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 70 at a dilution of 1:100-1:200 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunocytochemistry/Immunofluorescence: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Analysis of Heat Shock Protein 70 using Heat Shock Protein 70 Monoclonal antibody (3A3) shows staining in HeLa cells. Heat Shock Protein 70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 70 at a dilution of 1:100-1:200 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunohistochemistry-Paraffin: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Both normal and cancer biopsies of deparaffinized human Tonsil tissue.

Immunohistochemistry-Paraffin: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Both normal and cancer biopsies of deparaffinized human Testis tissue.

Immunoprecipitation: HSP70/HSPA1A Antibody (3A3) [NB600-1469] - Immunoprecipitation of Heat Shock Protein 70 (Hsp70) was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500ug whole cell lysate with 2ug of Hsp70 monoclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Plus Agarose at a dilution of 1:1000 overnight rotating at 4C, washed in TBST, and probed with goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:20,000 for at least 1 hour.

Product Discontinued

HSP70/HSPA1A Antibody (3A3) Summary

• Immunogen: Human recombinant HSP70 overexpressed in E. coli.
• Localization: Cytoplasmic
• Specificity: Epitope mapping with a panel of HSP70 deletion mutants suggests that the epitope recognized is located between amino acids 504-617 of human HSP70, a region that has been shown to be involved in stress-induced nucleolar localization.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: HSPA1A
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Gel Super Shift Assays 1:1 - 1:100
  • Immunocytochemistry/ Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation 2 ul
  • Western Blot 1:1000 - 1:5000
• Application Notes: WB: Detects an approx. proteins from ~70 kDa to ~78 kDa representing different members of the HSP70 family.

Reactivity Notes

Arthropod and Mollusca. Insect reactivity reported in scientific literature (PMID: 20798487). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Ascites diluted with PBS
• Preservative: 0.05% Sodium Azide
• Purity: Unpurified

Alternate Names for HSP70/HSPA1A Antibody (3A3)

• 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

The Hsp 70 family is a set of highly conserved proteins that are induced by a variety of biological stresses, including heat stress, in every organism in which the proteins have been examined. The human Hsp 70 family members include: Hsp 70, a protein which is strongly inducible in all organisms but which is also constitutively expressed in primate cells; Hsp 72, a 72 kDa protein that is induced exclusively under stress conditions; Hsc 70, or cognate protein, is a 72 kDa, constitutively expressed, protein which is involved in the uncoating of clathrin coated vesicles; GRP78, or BiP, is a glucose regulated 78 kDa protein localized in the endoplasmic reticulum; and p75, or Hsp 75, a 75 kDa protein that is found within the mitochondria.

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.

Publications for HSP70/HSPA1A Antibody (NB600-1469)(3)

Publications using NB600-1469

• Wang X, Duan C, Li Y et al. Sodium butyrate reduces overnutrition-induced microglial activation and hypothalamic inflammation International Immunopharmacology 2022-10-01 [PMID: 35917736] (WB, Mouse)
• Boussadia Z, Lamberti J, et al. Acidic microenvironment plays a key role in human melanoma progression through a sustained exosome mediated transfer of clinically relevant metastatic molecules. J Exp Clin Cancer Res 2018-10-05 [PMID: 30290833] (WB, Human)
• Blatzer M, Blum G, Jukic E et al. Blocking Hsp70 enhances the efficiency of Amphotericin B treatment in resistant Aspergillus terreus strains Antimicrob. Agents Chemother. 2015-04-13 [PMID: 25870060] (WB, Human)

Bioinformatics

• Gene Symbol: HSPA1A
• Entrez:
  • 24472(Rat)
• Uniprot:
  • P08107()