Fas antibody | knockout validation | Millipore 05-351

This is a knockout-validated antibody summary, based on the publication "The Fas/Fap-1/Cav-1 complex regulates IL-1RA secretion in mesenchymal stem cells to accelerate wound healing", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rat monoclonal IgG2a

Company: Millipore

Antibody: Fas

Catalog number: 05-351

Summary: Rat monoclonal IgG2a against a recombinant protein containing the cytoplasmic domain of mouse Fas. Reacts with mouse and bovine. Suitable for western blot and immunoprecipitation.

Validation Method

Western blot

Sample

Gingiva-derived Mesenchymal stem cells (GMSCs). Cells were lysed in M-PER mammalian protein extraction reagent (Thermo Fisher Scientific) with protease and phosphatase inhibitors (Roche).

Blocking agent

5% nonfat dry milk and 0.1% Tween 20 for 1 hour.

Primary incubation

Diluted in blocking solution overnight.

Secondary incubation

1:10,000 dilution horseradish peroxidase–conjugated secondary antibody (Santa Cruz Biotechnology in blocking solution for 1h at room temperature.

Detection

SuperSignal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific) and Biomax film (Kodak).

Figure

Western blotting of the indicated proteins from WT control and Fas-deficient MRL/lpr GMSCs (n = 3). Please see Figure 2e in the article [1].

References
  1. Kou X, Xu X, Chen C, Sanmillan M, Cai T, Zhou Y, et al. The Fas/Fap-1/Cav-1 complex regulates IL-1RA secretion in mesenchymal stem cells to accelerate wound healing. Sci Transl Med. 2018;10: pubmed publisher