Product nameAnti-HSF1 (phospho S303) antibody
See all HSF1 primary antibodies
DescriptionRabbit polyclonal to HSF1 (phospho S303)
Tested applicationsSuitable for: WB, IHC-P, ELISA, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Synthesized phosphopeptide derived from HSF1 around the phosphorylation site of serine 303 (Human).
- WB: MCF7 whole cell lysate (ab3871), Extracts from MCF cells. IHC-P: Human breast carcinoma tissue.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Without Mg+2 and Ca+2
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThe antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site.
Our Abpromise guarantee covers the use of ab47369 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Predicted molecular weight: 57 kDa.|
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
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
Lanes 1-2 : HSF1 antibody at 1/500 dilution
Lanes 3-4 : Anti-HSF1 (phospho S303) antibody (ab47369) at 1/500 dilution
Lanes 1 & 4 : Extracts from MCF cells treated with TNF alpha (20ng/ml, 30min). No peptide.
Lane 2 : Extracts from MCF cells treated with TNF alpha (20ng/ml, 30min). Synthetic phosphopeptide present.
Lane 3 : Extracts from MCF cells not treated
with TNF alpha. No peptide.
Predicted band size: 57 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?
ab47369 (1/50) staining HSF1 in paraffin-embedded human breast carcinoma tissue. Right-hand panel corresponds to a negative control for which ab47369 was pre-incubated with the immunising (blocking) peptide.
ICC/IF image of ab47369 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47369, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This product has been referenced in:
- Shao J et al. HSF1 phosphorylation by cyclosporin A confers hyperthermia sensitivity through suppression of HSP expression. Biochim Biophys Acta Gene Regul Mech N/A:N/A (2019). Read more (PubMed: 31054961) »
- Gomez-Pastor R et al. Abnormal degradation of the neuronal stress-protective transcription factor HSF1 in Huntington's disease. Nat Commun 8:14405 (2017). WB, IHC . Read more (PubMed: 28194040) »