Expression and Purification of Human Heat-Shock Transcription Factor 1
Copyright policy )Expression and Purification of Human Heat-Shock Transcription Factor 1
Heat-shock factor 1 (HSF1) is a 57-kDa cytoplasmic protein which binds to the promoters of heat-shock genes and activates transcription during heat shock. We describe here the expression and purification of the 529 amino acids form of human HSF1. We designed a new and complete purification protocol involving ammonium sulfate precipitation, heparin-Sepharose affinity, and ion-exchange chromatography, which allows the purification of large amounts of pure and active recombinant protein. HSF1 isolated by this method is pure as that assessed by SDS-PAGE and reverse-phase HPLC. The purified protein is recognized by specific anti-HSF1 antibodies, binds to heat-shock elements, and activates the promoter of the heat-shock protein 70A gene in vitro.
- Inserm France
- Dana-Farber Cancer Institute United States
- Harvard University United States
Transcription, Genetic, Protein Conformation, Regulatory Sequences, Nucleic Acid, Recombinant Proteins, DNA-Binding Proteins, [SDV.CAN] Life Sciences [q-bio]/Cancer, Gene Expression Regulation, Heat Shock Transcription Factors, Humans, HSP70 Heat-Shock Proteins, Cloning, Molecular, Heat-Shock Proteins, Protein Binding, Transcription Factors
Transcription, Genetic, Protein Conformation, Regulatory Sequences, Nucleic Acid, Recombinant Proteins, DNA-Binding Proteins, [SDV.CAN] Life Sciences [q-bio]/Cancer, Gene Expression Regulation, Heat Shock Transcription Factors, Humans, HSP70 Heat-Shock Proteins, Cloning, Molecular, Heat-Shock Proteins, Protein Binding, Transcription Factors
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