HSF1 antibody | knockout validation | Cell Signaling 12972

This is a knockout-validated antibody summary, based on the publication "HSF1 stress response pathway regulates autophagy receptor SQSTM1/p62-associated proteostasis", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: HSF1

Catalog number: 12972

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxy terminus of human HSF1 protein. Reacts with human, mouse, rat, monkey, bovine, dog, and pig. Suitable for western blot, immunoprecipitation, immunohistochemistry (paraffin), immunofluorescence/immunocytochemistry, flow cytometry and ChIP.

Validation Method

Western blot

Sample

WT and HSF1-KO HeLa cells. Cells were extracted with ice-cold PBS containing 0.5% Triton X-100 (Wako Pure Chemical Industries, 168-11805) and a protease inhibitor cocktail (Nacalai Tesque, 25955-11).

Blocking agent

10% skimmed milk, 5% bovine serum albumin (Nacalai Tesque, 01860-36), or Blocking Solution (Nacalai Tesque, 05151-35) for 1 h.

Primary incubation

For 12 h at 25°C.

Secondary incubation

1:4,000 dilution alkaline phosphatase-conjugated anti-rabbit IgG (Jackson ImmunoResearch Laboratories Inc., 111-055-144).

Detection

Nitroblue tetrazolium chloride (Nacalai Tesque, 24720-01) and 5-bromo-4-chloro-3-indolylphosphate P-toluidine salt (Nacalai Tesque, 05643-11) reagents.

Figure

Cell lysates of WT and individual HSF1-KO HeLa cell clones (#6 and #12) were subjected to immunoblot analysis with anti-HSF1 and anti-actin. From [1].

References
  1. Watanabe Y, Tsujimura A, Taguchi K, Tanaka M. HSF1 stress response pathway regulates autophagy receptor SQSTM1/p62-associated proteostasis. Autophagy. 2017;13:133-148 pubmed publisher