TRC8 antibody | knockout validation | Santa Cruz sc-390347

This is a knockout-validated antibody summary, based on the publication "Insulin-induced gene 1 (INSIG1) inhibits HIV-1 production by degrading Gag via activity of the ubiquitin ligase TRC8", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Mouse monoclonal IgG2a (kappa light chain)

Company: Santa Cruz

Antibody: TRC8

Catalog number: sc-390347

Summary: Mouse monoclonal IgG2a (kappa light chain) against an internal region of human TRC8 (amino acids 106-124). Recommended for detection of TRC8 of mouse, rat and human origin by western blot, immunoprecipitation, immunofluorescence and ELISA. Also reactive with additional species, including equine, canine, bovine and porcine.

Validation Method

Western blot

Sample

Control and Trc8-knockout 293T cells. Cells were lysed in NP-40 buffer (Nonidet P-40, 20 mM Tris HCl pH 8.0, 137 mM NaCl, 10% glycerol, 2 mM EDTA) for 30 min on ice for lysis.

Blocking agent

5% skim milk.

Secondary incubation

Goat anti-mouse IgG-HRP (Sungene Biotech).

Detection

Tannon-5500 gel imager.

Figure

Western blot of the indicated proteins in Amfr knockout, Trc8 knockout and control 293T cells co-transfected with INSIG1 and Gag-pol plasmids. Please see Figure 7a in the article [1].

References
  1. Zhang Y, Lu J, Ma J, Liu X. Insulin-induced gene 1 (INSIG1) inhibits HIV-1 production by degrading Gag via activity of the ubiquitin ligase TRC8. J Biol Chem. 2019;294:2046-2059 pubmed publisher