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  1. Link

    hsf1-antibody-ep1710y-chip-grade-ab52757.pdf

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Tags & Cell Markers Subcellular Markers Nucleus Other Nuclear Bodies
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

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  • SDS
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Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunoprecipitation - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • ChIP - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Flow Cytometry (Intracellular) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Flow Cytometry (Intracellular) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
  • Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1710Y] to HSF1 - ChIP Grade
  • Suitable for: Flow Cyt (Intra), ChIP, WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free FITC HRP PE

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Overview

  • Product name

    Anti-HSF1 antibody [EP1710Y] - ChIP Grade
    See all HSF1 primary antibodies
  • Description

    Rabbit monoclonal [EP1710Y] to HSF1 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ChIP, WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human HSF1 aa 450 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: K562 HAP1 and HeLa whole cell lysate (ab150035). ICC/IF: MCF-7 cells. Flow Cyt (intra): HeLa cells. IHC-P: Human ovarian carcinoma tissue; Mouse testis and colon tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.21% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1710Y
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Other Nuclear Bodies
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Anti-HSF1 antibody [EP1710Y] - BSA and Azide free (ab232342)
    • FITC Anti-HSF1 antibody [EP1710Y] (ab237150)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human HSF1 protein (ab78795)
  • Related Products

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52757 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 (Intra)
1/20.

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

ChIP
Use at an assay dependent concentration.
WB
1/50000. Detects a band of approximately 85 kDa (predicted molecular weight: 57 kDa).

For unpurified use at 1/100000. 

IP
1/20.

For unpurified use at 1/100.

IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
Notes
Flow Cyt (Intra)
1/20.

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

ChIP
Use at an assay dependent concentration.
WB
1/50000. Detects a band of approximately 85 kDa (predicted molecular weight: 57 kDa).

For unpurified use at 1/100000. 

IP
1/20.

For unpurified use at 1/100.

IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.

Target

  • Function

    DNA-binding protein that specifically binds heat shock promoter elements (HSE) and activates transcription. In higher eukaryotes, HSF is unable to bind to the HSE unless the cells are heat shocked.
  • Sequence similarities

    Belongs to the HSF family.
  • Domain

    the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
  • Post-translational
    modifications

    Phosphorylated on multiple serine residues, a subset of which are involved in stress-related regulation of transcription activation. Constitutive phosphorylation represses transcriptional activity at normal temperatures. Levels increase on specific residues heat-shock and enhance HSF1 transactivation activity. Phosphorylation on Ser-307 derepresses activation on heat-stress and in combination with Ser-303 phosphorylation appears to be involved in recovery after heat-stress. Phosphorylated on Ser-230 by CAMK2, in vitro. Cadmium also enhances phosphorylation at this site. Phosphorylation on Ser-303 is a prerequisite for HSF1 sumoylation. Phosphorylation on Ser-121 inhibits transactivation and promotes HSP90 binding. Phosphorylation on Thr-142 also mediates transcriptional activity induced by heat. Phosphorylation on Ser-326 plays an important role in heat activation of HSF1 transcriptional activity.
    Sumoylated with SUMO1 and SUMO2 on heat-shock. Heat-inducible sumoylation occurs after 15 min of heat-shock, after which levels decrease and at 4 hours, levels return to control levels. Sumoylation has no effect on HSE binding nor on transcriptional activity. Phosphorylation on Ser-303 is a prerequisite for sumoylation.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasmic during normal growth. On activation, translocates to nuclear stress granules. Colocalizes with SUMO1 in nuclear stress granules.
  • Target information above from: UniProt accession Q00613 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3297 Human
    • Entrez Gene: 15499 Mouse
    • Omim: 140580 Human
    • SwissProt: Q00613 Human
    • SwissProt: P38532 Mouse
    • Unigene: 530227 Human
    • Unigene: 347444 Mouse
    • Alternative names

      • Heat shock factor 1 antibody
      • Heat shock factor protein 1 antibody
      • Heat shock transcription factor 1 antibody
      • HSF 1 antibody
      • hsf1 antibody
      • HSF1_HUMAN antibody
      • HSTF 1 antibody
      • HSTF1 antibody
      see all

    Images

    • Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      All lanes : Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757) at 1/100000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : Hsf1 knockout HAP1 whole cell lysate
      Lane 3 : Hela whole cell lysate
      Lane 4 : K562 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 57 kDa
      Observed band size: 57 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab52757 observed at 57 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab52757 was shown to specifically react with in wild-type HAP1 cells as signal was lost in Hsf1 knockout cells. Wild-type and Hsf1 knockout samples were subjected to SDS-PAGE. Ab52757 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/100000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human ovarian carcinoma tissue sections labeling HSF1 with Purified ab52757 at 1:250 dilution (1.06 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • Immunoprecipitation - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunoprecipitation - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      ab52757 (purified) at 1:20 dilution (2μg) immunoprecipitating HSF1 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
      Lane 2 (+): ab52757 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52727 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • ChIP - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      ChIP - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Chromatin was prepared from HeLa cells heat shocked (42oC 30 minutes) according to the Abcam Dual X-ChIP protocol. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
      The ChIP was performed with 25 µg of chromatin, 5 µg of ab52757 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).
      Primers and probes are located in the first kb of the transcribed region.
      *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling HSF1 with Purified ab52757 at 1:100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry (Intracellular) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Flow Cytometry (Intracellular) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HSF1 with purified ab52757 at 1/20 dilution (10µg/ml) (red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

       

       

    • Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757) at 1/50000 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell ) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 57 kDa
      Observed band size: 80 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer: 5% NFDM/TBST

    • Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Western blot - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757) at 1/100000 dilution (unpurified) + HeLa cell lysate at 10 µg

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 57 kDa
      Observed band size: 85 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling HSF1 with ab52757 at 1/1000 followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Nuclear staining on mouse testis. The section was incubated with ab52757 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), Ready to use.

      Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling HSF1 with ab52757 at 1/1000 followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Nuclear staining on mouse colon. The section was incubated with ab52757 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), Ready to use.

      Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      ab52757 staining HSF1 in wild-type Hap1 cells (top panel) and HSF1 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab52757 at 1/250 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunocytochemistry/ Immunofluorescence - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling HSF1 with unpurified ab52757 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
      Control: PBS only.
      Nuclear counter stain: DAPI.

    • Flow Cytometry (Intracellular) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Flow Cytometry (Intracellular) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Overlay histogram showing HeLa cells stained withunpurified ab52757 (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 (unpurified ab52757, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. 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. 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

      Immunohistochemical staining of paraffin-embedded human ovarian carcinoma using unpurified ab52757 at a 1:100 dilution.

    • Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)
      Anti-HSF1 antibody [EP1710Y] - ChIP Grade (ab52757)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (24)

    Publishing research using ab52757? Please let us know so that we can cite the reference in this datasheet.

    ab52757 has been referenced in 24 publications.

    • Chen F  et al. Combination of HSP90 and autophagy inhibitors promotes hepatocellular carcinoma apoptosis following incomplete thermal ablation. Mol Med Rep 22:337-343 (2020). PubMed: 32319654
    • Mun GI  et al. Decreased expression of FBXW7 by ERK1/2 activation in drug-resistant cancer cells confers transcriptional activation of MDR1 by suppression of ubiquitin degradation of HSF1. Cell Death Dis 11:395 (2020). PubMed: 32457290
    • Song P  et al. ß-catenin represses miR455-3p to stimulate m6A modification of HSF1 mRNA and promote its translation in colorectal cancer. Mol Cancer 19:129 (2020). PubMed: 32838807
    • Zhou G  et al. Role of the Receptor for Advanced Glycation End Products in Heat Stress-Induced Endothelial Hyperpermeability in Acute Lung Injury. Front Physiol 11:1087 (2020). PubMed: 33192536
    • Su KH  et al. Heat Shock Factor 1 Is a Direct Antagonist of AMP-Activated Protein Kinase. Mol Cell 76:546-561.e8 (2019). PubMed: 31561952
    View all Publications for this product

    Customer reviews and Q&As

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    Please use the links above to contact us or submit feedback about this product.

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