Overview

  • Product nameAnti-Hsc70 antibody [13D3]See all Hsc70 primary antibodies ...
  • Description
    Mouse monoclonal [13D3] to Hsc70
  • Specificityab2788 detects heat shock cognate protein 70 kDa (HSC70) from human, feline, primate, rat, and mouse tissues. This antibody displays slight cross-reactivity to HSP70.
  • Tested applicationsICC/IF, Inhibition Assay, IHC-Fr, IP, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Cow, Cat, Human, Non Human Primates
    Predicted to work with: Horse, Chicken, Fish
  • Immunogen

    Full length protein corresponding to Mouse Hsc70. Mouse spermatogenic cell protein
    Database link: P11142

  • General notes

    HSC70 (also known as HSC71, HSC73, HSP73, p72, prp73) is expressed constitutively and is slightly heat-inducible. HSC70 binds to the exposed loop of clathrin light chains to promote uncoating and can also bind the cytoskeleton which may facilitate cytoskeletal rearrangements. HSC70 has been shown to stimulate lysosomal degradation of intracellular proteins and to retard both aggregation and folding of mitochondrial precursor proteins in vitro.

     

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 bufferPreservative: 0.05% Sodium azide
    Constituent: PBS
  • 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; GRP 78, or BiP, is a glucose regulated 78 kDa protein localized in the endoplasmic reticulum; and p75, or Hsc 75, a 75 kDa protein that is found within the mitochondria. Hsc 70 (also known as Hsc 71, Hsc 73, Hsc 73, p72, prp 73) is expressed constitutively and is slightly heat-inducible. Hsc 70 binds to the exposed loop of clathrin light chains to promote uncoating and can also bind the cytoskeleton which may facilitate cytoskeletal rearrangements. Hsc 70 has been shown to stimulate lysosomal degradation of intracellular proteins and to retard both aggregation and folding of mitochondrial precursor proteins in vitro.
  • Clonality Monoclonal
  • Clone number13D3
  • IsotypeIgM
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab2788 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
ICC/IF 1/20 - 1/200.
Inhibition Assay Use at an assay dependent concentration.
IHC-Fr 1/20 - 1/200.
IP Use at an assay dependent concentration.

 Suggest 2 μl

WB 1/1000 - 1/5000. Detects a band of approximately 70 kDa.
Flow Cyt 1/200.

Target

  • FunctionActs as a repressor of transcriptional activation. Inhibits the transcriptional coactivator activity of CITED1 on Smad-mediated transcription. Chaperone. Isoform 2 may function as an endogenous inhibitory regulator of HSC70 by competing the co-chaperones.
  • Tissue specificityUbiquitous.
  • Sequence similaritiesBelongs to the heat shock protein 70 family.
  • DomainThe N-terminal 1-386 residues constitute the ATPase domain, while residues 387-646 form the peptide-binding domain.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
    ISGylated.
  • Cellular localizationCytoplasm. Melanosome. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Translocates rapidly from the cytoplasm to the nuclei, and especially to the nucleoli, upon heat shock. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2410008N15Rik antibody
    • Constitutive heat shock protein 70 antibody
    • Heat shock 70 kDa protein 8 antibody
    • Heat shock 70kD protein 10 antibody
    • Heat shock 70kD protein 8 antibody
    • Heat shock 70kDa protein 8 antibody
    • Heat shock cognate 71 kDa protein antibody
    • Heat shock cognate 71 kDa protein. antibody
    • Heat shock cognate protein 54 antibody
    • Heat shock cognate protein 71 kDa antibody
    • Heat shock cognate protein 71-kDa antibody
    • Heat shock protein 8 antibody
    • Heat shock protein A8 antibody
    • Heat-shock70-KD protein 10, formerly antibody
    • HSC54 antibody
    • HSC71 antibody
    • Hsc73 antibody
    • HSP71 antibody
    • HSP73 antibody
    • HSP7C_HUMAN antibody
    • HSPA10 antibody
    • HSPA8 antibody
    • LAP1 antibody
    • Lipopolysaccharide associated protein 1 antibody
    • LPS associated protein 1 antibody
    • LPS associated protein antibody
    • MGC102007 antibody
    • MGC106514 antibody
    • MGC114311 antibody
    • MGC118485 antibody
    • MGC131511 antibody
    • MGC29929 antibody
    • N-myristoyltransferase inhibitor protein 71 antibody
    • NIP71 antibody
    see all

Anti-Hsc70 antibody [13D3] images

  • Immunohistochemistry was performed on both normal and 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 1:200 with a mouse monoclonal antibody recognizing Heat Shock Cognate Protein 70 (ab2788) 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.
  • Immunocytochemistry/Immunofluorescence analysis of Hsc70 (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% BSA for 15 minutes at room temperature. Cells were incubated with ab2788 (1:50) for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488 goat anti-mouse IgG secondary antibody (1:400) for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

  • Western blot analysis of Hsc70 was performed by loading 50µg of the indicated whole cell lysates and 15µl of prestained protein ladder onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was incubated with ab2788 (1:1000) overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and incubated with a goat anti-mouse IgM-HRP secondary antibody (1:20,000) for at least 1 hour. Chemiluminescent detection was performed.

  • Immunohistochemistry was performed on both normal and cancer 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 Cognate Protein 70 (ab2788) 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.
  • ab2788 (2µ/ml) staining Hsc70in human frontal cortex using an automated system. Using this protocol there is cytoplasmic staining of nerons. Inset shows non-immune IgG incubation.
    Sections were rehydrated and antigen retrieved with a buffer containing EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with an amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human Testis 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 Cognate Protein 70 (ab2788) 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.
  • Immunofluorescent analysis of Heat Shock Protein 70 using Heat Shock Protein 70 Monoclonal antibody (13D3) ab2788 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 ab2788 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of Heat Shock Protein 70 using Heat Shock Protein 70 Monoclonal antibody (13D3) ab2788 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 ab2788 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of Heat Shock Protein 70 using Heat Shock Protein 70 Monoclonal antibody (13D3) ab2788 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. 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 ab2788 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Overlay histogram showing HeLa cells stained with ab2788 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2788, 1/200 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-Hsc70 antibody [13D3] (ab2788)

This product has been referenced in:
  • Zhang L  et al. HACE1-dependent protein degradation provides cardiac protection in response to haemodynamic stress. Nat Commun 5:3430 (2014). WB ; Mouse . Read more (PubMed: 24614889) »
  • Fernandez-Estevez MA  et al. Trehalose reverses cell malfunction in fibroblasts from normal and Huntington's disease patients caused by proteosome inhibition. PLoS One 9:e90202 (2014). ICC ; Human . Read more (PubMed: 24587280) »

See all 6 Publications for this product

Product Wall

Application Western blot
Sample Mouse Tissue lysate - whole (liver)
Loading amount 15 µg
Specification liver
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Sep 05 2012

Thank you for your enquiry. I am sorry to hear that your customer has been having difficulties with this antibody by immunocytochemistry. From a literature search I have determined that this heat shock protein is largely cytoplasmic. For example in...

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Application Western blot
Sample Mouse Tissue lysate - whole (liver)
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Oct 20 2005

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soona s any new information is brought to our attention. As far as we are aware, cross reactivity with Drosoph...

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