HSF1 Antibody

Western Blot: HSF1 Antibody [NB300-730] - Analysis of Heat Shock Factor 1 (HSF1) was performed by loading 50ug 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 probed with a HSF1 rabbit polyclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:20,000 for at least one hour. Chemiluminescent detection was performed using SuperSignal West Pico.

Immunocytochemistry/Immunofluorescence: HSF1 Antibody [NB300-730] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a HSF1 polyclonal antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.

Immunoprecipitation: HSF1 Antibody [NB300-730] - Analysis of Heat Shock Factor 1 (HSF 1) was performed on HeLa cells. Antigen: antibody complexes were formed by incubating 500ug whole cell lysate with 2ug of HSF1 polyclonal antibody overnight on a rocking platform at 4C. Immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with 5X Lane Marker Reducing Sample 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 probed with a HSF1 polyclonal antibody at a dilution of 1:1000 overnight rotating at 4C, washed in TBST, and probed with Clean Blot IP Detection Reagent-HRP at a dilution of 1:1000 for at least one hour. Chemiluminescent detection was performed using SuperSignal West Dura.

Product Discontinued

HSF1 Antibody Summary

• Immunogen: Recombinant human HSF1 expressed in E. coli.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HSF1
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100 - 1:2000
  • Gel Super Shift Assays 1:1 - 1:100
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Immunoprecipitation 2 ug
  • Western Blot 1:1000 - 1:10000
• Application Notes: WB: Detects an approx. 83 kDa protein representing HSF1 from NIH/3T3 cell lysate. IP: Detects an approx. 69 kDa protein representing HSF1 from HelaS3 cells.
• Control:

  NIH 3T3 Whole Cell Lysate

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Whole antisera with PBS
• Preservative: 0.05% Sodium Azide
• Purity: Unpurified

Alternate Names for HSF1 Antibody

• heat shock factor protein 1
• heat shock transcription factor 1HSTF1HSF 1
• HSF1
• HSTF 1
• HSTF1

Background

All organisms respond to elevated temperatures and a variety of environmental stresses by rapid synthesis of heat shock RNAs and proteins. The regulation of heat shock gene transcription is mediated by the transcriptional activator, heat shock factor (HSF), which binds to heat shock response elements (HSEs). These HSEs are found as three repeats of a 5-nucleotide {nGAAn} module, arranged in alternating orientation and present upstream of all heat shock genes. The HSEs are highly conserved among species yet HSF purified from yeast, Drosophila and human have different molecular weights and the proteins do not show significant immunological cross reaction. Two HSFs have been identified in human cells, HSF 1 and HSF 2, which bind to the same HSEs and have 38% sequence identity. These factors are activated by distinct stimuli, HSF 1 is responsive to classical stress signals such as heat, heavy metals and oxidative reagents, whereas HSF 2 is activated during hemin-mediated differentiation of human erythroleukemia cells. HSF 1 exists constitutively in the cytoplasm and the nucleus of unstressed cells as a monomer which lacks DNA binding activity. Through an unknown signal generated during stress, HSF 1 becomes activated to a nuclear localized, trimeric state which binds to DNA. The phosphorylation of HSF 1 is necessary for maximal transcription of heat shock genes.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HSF1 Antibody (NB300-730)(1)

Publication using NB300-730

• Rogers RS, Morris EM, Wheatley JL et al. Deficiency in the Heat Stress Response Could Underlie Susceptibility to Metabolic Disease. Diabetes. 2016-08-23 [PMID: 27554472] (WB, Rat)

Bioinformatics

• Gene Symbol: HSF1
• Entrez:
  • 3297(Human)
  • 15499(Mouse)
  • 79245(Rat)