Overview

  • Product nameAnti-Hsc70 antibody [EP1531Y]
    See all Hsc70 primary antibodies
  • Description
    Rabbit monoclonal [EP1531Y] to Hsc70
  • Tested applicationsSuitable for: Flow Cyt, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Hsc70 aa 600 to the C-terminus (C terminal).

  • Positive control
    • WB: HeLa cell lysate IHC-P: Human breast carcinoma tissue ICC/IF: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-Hsc70 antibody (HRP) [EP1531Y] (ab200757)

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • PurityProtein A purified
  • ClonalityMonoclonal
  • Clone numberEP1531Y
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab51052 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
Flow Cyt 1/30 - 1/70.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/10 - 1/20.
WB 1/500 - 1/5000. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/100 - 1/250.

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
    • Epididymis luminal protein 33 antibody
    • Epididymis secretory sperm binding protein Li 72p 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 protein 54 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
    • HEL 33 antibody
    • HEL S 72p 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 [EP1531Y] images

  • All lanes : Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/5000 dilution (purified)

    Lane 1 : C6 cell lysate
    Lane 2 : PC-12 cell lysate
    Lane 3 : mouse heart lysate
    Lane 4 : rat heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 71 kDa
    Observed band size : 71 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/5000 dilution (purified)

    Lane 1 : human fetal brain lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : HEK293 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 71 kDa
    Observed band size : 71 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of MCF7 cells with purified ab51052 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab51052 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab51052 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • ab51052 (purified) at 1/20 immunoprecipitating Hsc70 in 10 μg HeLa (Lanes 1 and 2, observed at 71 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Overlay histogram showing MCF7 cells fixed in 4% PFA and stained with purified ab51052 at a dilution of 1 in 70 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  • Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/500 dilution (unpurified) + HeLa cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled at 1/2000 dilution

    Predicted band size : 71 kDa
    Observed band size : 71 kDa
  • Unpurified ab51052 (1/250) staining Hsc70 in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.

    Ab51052 (1/250) staining Hsc70 in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.
  • Immunofluorescent staining of HeLa cells using unpurified ab51052 (1/100).

References for Anti-Hsc70 antibody [EP1531Y] (ab51052)

This product has been referenced in:
  • Jakobsson ME  et al. Identification and characterization of a novel human methyltransferase modulating Hsp70 protein function through lysine methylation. J Biol Chem 288:27752-63 (2013). WB . Read more (PubMed: 23921388) »
  • Leu JI  et al. HSP70 Inhibition by the Small-Molecule 2-Phenylethynesulfonamide Impairs Protein Clearance Pathways in Tumor Cells. Mol Cancer Res 9:936-47 (2011). Human . Read more (PubMed: 21636681) »

See all 4 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Hamster Cell lysate - whole cell (CHO cells)
Gel Running Conditions Reduced Denaturing
Loading amount 30 µg
Specification CHO cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Sep 05 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (MEFs)
Permeabilization Yes - 1% triton x-100
Specification MEFs
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: rt°C
Fixative Paraformaldehyde
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Submitted Sep 02 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HEK293)
Total protein in input 200 µg
Immuno-precipitation step Other - Dynabeads IgG
Specification HEK293
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Submitted Feb 08 2016

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Chinese Hamster Cell lysate - whole cell (CHO cell)
Specification CHO cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Jan 09 2015

Application Immunoprecipitation
Immuno-precipitation step Other - IgG Dynabead
Sample Mouse Tissue lysate - whole (mouse heart)
Specification mouse heart
Total protein in input 200 µg
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Submitted Oct 14 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Oct 09 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (h9c2)
Specification h9c2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Oct 09 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (mouse heart)
Specification mouse heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Oct 09 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Mouse Cell (primary cortical neurons)
Specification primary cortical neurons
Permeabilization Yes - 0.1% Triton/PBS
Fixative Paraformaldehyde
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Submitted Jun 23 2014

Application Immunoprecipitation
Immuno-precipitation step Other - Protein G Dynabeads crosslinked to antibody
Sample Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
Specification mouse embryonic fibroblasts
Total protein in input 500 µg
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Submitted Jun 09 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"