Overview

  • Product nameAnti-Hsp70 antibody [4G4]See all Hsp70 primary antibodies ...
  • Description
    Mouse monoclonal [4G4] to Hsp70
  • Specificityab5444 detects heat shock protein 70 kDa (Hsp 70) from human tissues. This antibody specifically detects Hsp 70 and does not cross-react with other members of the Hsp 70 family.
  • Tested applicationsInhibition Assay, IHC-P, ICC/IF, Flow Cyt, Blocking, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Dog, Pig, Non Human Primates
  • Immunogen

    Recombinant fragment corresponding to Human Hsp70.

  • EpitopeEpitope 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.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferConstituents: 99% PBS, Ascites
  • Concentration information loading...
  • PurityAscites
  • Primary antibody notes 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.
  • ClonalityMonoclonal
  • Clone number4G4
  • IsotypeIgG1
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab5444 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Inhibition Assay Use at an assay dependent concentration.
IHC-P 1/20 - 1/50.
ICC/IF 1/100.
Flow Cyt 1/100 - 1/200.
Blocking Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Detects a band of approximately 70 kDa.

Target

  • FunctionIn cooperation with other chaperones, Hsp70s stabilize preexistent proteins against aggregation and mediate the folding of newly translated polypeptides in the cytosol as well as within organelles. These chaperones participate in all these processes through their ability to recognize nonnative conformations of other proteins. They bind extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In case of rotavirus A infection, serves as a post-attachment receptor for the virus to facilitate entry into the cell.
  • Tissue specificityHSPA1B is testis-specific.
  • Sequence similaritiesBelongs to the heat shock protein 70 family.
  • Cellular localizationCytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt
  • Database links
  • Alternative names
    • DAQB 147D11.1 001 antibody
    • DnaK type molecular chaperone HSP70 1 antibody
    • Epididymis secretory protein Li 103 antibody
    • FLJ54303 antibody
    • FLJ54370 antibody
    • FLJ54392 antibody
    • FLJ54408 antibody
    • FLJ75127 antibody
    • Heat shock 70 kDa protein 1 antibody
    • Heat shock 70 kDa protein 1/2 antibody
    • Heat shock 70 kDa protein 1A/1B antibody
    • heat shock 70kDa protein 1A antibody
    • Heat shock 70kDa protein 1B antibody
    • Heat shock induced protein antibody
    • heat shock protein 70 antibody
    • HEL S 103 antibody
    • HSP70 1 antibody
    • HSP70 1B antibody
    • HSP70 2 antibody
    • HSP70-1/HSP70-2 antibody
    • HSP70-1A antibody
    • HSP70.1 antibody
    • HSP70.1/HSP70.2 antibody
    • HSP70I antibody
    • HSP71_HUMAN antibody
    • HSP72 antibody
    • HSPA1 antibody
    • HSPA1A antibody
    • HSPA1B antibody
    • XXbac BCX40G17.3 001 antibody
    see all

Anti-Hsp70 antibody [4G4] images

  • Immunocytochemistry/Immunofluorescence analysis of Hsp70 in HeLa Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab5444 at a dilution of 1:100 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Hsp70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp70 in NCI-H1299 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab5444 at a dilution of 1:100 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Hsp70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp70 in NIH-3T3 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab5444 at a dilution of 1:100 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Hsp70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunoprecipitation of Hsp70 was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500ug whole cell lysate with 2ug of Hsp70 monoclonal antibody (ab5444) overnight on a rocking platform at 4�C. The immune complexes were captured on 50ul Protein Agarose washed extensivelyand eluted with Buffer. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel then transferred to a PVDF membraneand blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with Hsp70 monoclonal antibody (ab5444) at a dilution of 1:1000 overnight rotating at 4�C then washed in TBST and probed with Detection Reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed.
  • All lanes : Anti-Hsp70 antibody [4G4] (ab5444) at 1/1000 dilution

    Lane 1 : 293T cell lysates at 50 µg
    Lane 2 : HeLa cell lysates at 50 µg
    Lane 3 : K562 cell lysates at 50 µg
    Lane 4 : A431 cell lysates at 50
    Lane 5 : HepG2 cell lysates at 50

    Secondary
    HRP-conjugated goat anti-mouse IgG at 1/20000 dilution
  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human breast tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a mouse monoclonal antibody recognizing Heat Shock Protein 70 ab5444 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on cancer biopsies of deparaffinized Human prostate carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Heat Shock Protein 70 ab5444 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human tonsil tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Heat Shock Protein 70 ab5444 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References for Anti-Hsp70 antibody [4G4] (ab5444)

This product has been referenced in:
  • Heldens L  et al. An atypical unfolded protein response in heat shocked cells. PLoS One 6:e23512 (2011). WB ; Human . Read more (PubMed: 21853144) »
  • Marozkina NV  et al. Hsp 70/Hsp 90 organizing protein as a nitrosylation target in cystic fibrosis therapy. Proc Natl Acad Sci U S A 107:11393-8 (2010). WB ; Human . Read more (PubMed: 20534503) »

See all 3 Publications for this product

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