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Other Immunogen Type corresponding to Human HSF1. Recombinant human HSF1 expressed in E. coli.
Our Abpromise guarantee covers the use of ab2923 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|EMSA||Use at an assay dependent concentration. PubMed: 18429957|
|Gel supershift assays||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.
|WB||1/1000 - 1/10000. Detects a band of approximately 83 kDa (predicted molecular weight: 57 kDa).|
|ELISA||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
Immunocytochemistry/Immunofluorescence analysis of HSF1 (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 probed with ab2923 (1:50) for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat-anti rabbit 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.
Immunoprecipitation of HSF1 was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500µg whole cell lysate with 2µg of ab2923 overnight on a rocking platform at 4°C. Immune complexes were captured on 50µl Protein A/G Plus Agarose, washed extensively, and eluted with buffer. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was incubated with ab2923 (1:1000) overnight rotating at 4°C, washed in TBST, and probed with detection reagent (1:1000) for at least one hour. Chemiluminescent detection was performed.
Western blot analysis of Heat Shock Factor 1 (HSF1) was performed by loading 50µg of the indicated whole cell lysates per well 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 ab2923 (1/1000) overnight at 4°C on a rocking platform, washed in TBS-0.1% Tween 20, and probed with a HRP-conjugated goat anti-rabbit IgG secondary antibody (1/20,000) for at least one hour. Chemiluminescent detection was performed.
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