GP78-1 antibody | knockout validation | Santa Cruz sc-166358

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 IgG1 (kappa light chain)

Company: Santa Cruz

Antibody: GP78-1

Catalog number: sc-166358

Summary: Mouse monoclonal IgG1 (kappa light chain) against an internal region of human GP78-1 (AMFR, amino acids 87-116). Recommended for detection of GP78 isoform 1 of mouse, rat and human origin by western blot, immunoprecipitation, immunofluorescence and ELISA.

Validation Method

Western blot

Sample

Control and Amfr-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