Product nameAnti-HSF1 antibody [10H8]
See all HSF1 primary antibodies
DescriptionRat monoclonal [10H8] to HSF1
Tested applicationsSuitable for: IHC-P, Flow Cyt, ICC/IF, WB, IP, ICC, ELISAmore details
Species reactivityReacts with: Mouse, Rat, Rabbit, Guinea pig, Hamster, Cow, Human, Monkey
Purified recombinant HSF1 protein
- HeLa Cell Lysate This antibody gave a positive result in IHC in the following FFPE tissue: Human normal testis.
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: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.4
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab61382 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|EMSA||Use at an assay dependent concentration.|
|ICC/IF||1/100 - 1/200.|
|WB||1/1000. Predicted molecular weight: 57 kDa.|
|IP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
FunctionDNA-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 similaritiesBelongs to the HSF family.
Domainthe 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
modificationsPhosphorylated 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 localizationCytoplasm. Nucleus. Cytoplasmic during normal growth. On activation, translocates to nuclear stress granules. Colocalizes with SUMO1 in nuclear stress granules.
- Information by UniProt
- Heat shock factor 1 antibody
- Heat shock factor protein 1 antibody
- Heat shock transcription factor 1 antibody
All lanes : Anti-HSF1 antibody [10H8] (ab61382) at 1/1000 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
Lanes 1 - 4: Merged signal (red and green). Green - ab61382 observed at 57 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab61382 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in Hsf1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Hsf1 knockout samples were subjected to SDS-PAGE. Ab61382 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rat IgG H&L (IRDye® 800CW) preabsorbed and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Heat shocked, 2% formaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma)cells stained for HSF1 (green) using ab61382 at 1/100 dilution in ICC/IF. The nuclear counter stain is DAP (blue) The right hand panel shows the merged images.
Overlay histogram showing HeLa cells stained with ab61382 (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 (ab61382, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG1 [RTK2071] (ab18412, 1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). 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.
IHC image of HSF1 staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab61382, 5µg/ml, for 15 mins at room temperature. A Goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
This product has been referenced in:
- Dai F et al. Downregulation of MiR-199b-5p Inducing Differentiation of Bone-Marrow Mesenchymal Stem Cells (BMSCs) Toward Cardiomyocyte-Like Cells via HSF1/HSP70 Pathway. Med Sci Monit 24:2700-2710 (2018). Read more (PubMed: 29715263) »
- Chooi WH et al. Loading-Induced Heat-Shock Response in Bovine Intervertebral Disc Organ Culture. PLoS One 11:e0161615 (2016). IF . Read more (PubMed: 27580124) »