This is a knockout-validated antibody summary, based on the publication "Mechanical regulation of glycolysis via cytoskeleton architecture", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signalling

Antibody: TRIM21

Catalog number: 92043

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Val309 of human TRIM21 protein. Reacts with human, by western blot and immunoprecipitation.

Western blot

Control and TRIM21-KO NSCLC cells.

5% bovine serum albumin (BSA, BAH65–0500, Equitech-Bio, Kerrville, TX) in Tris buffered saline (TBS) with 0.1% Tween 20 (TBST, BP337, Fisher) for 1 hr at room temperature.

5% BSA in TBST for overnight at 4°C.

Secondary antibodies conjugated with horseradish peroxidase (goat anti rabbit: 111–035-003, Jackson ImmunoResearch, West Grove, PA).

Enhanced chemiluminescence using freshly prepared substrate solution based on 100 mM Tris (Tris-base, BP152 Fisher Scientific) adjusted to pH 8.5 with Hydrochloric acid (A144S-212, Fisher), 1.2 mM Luminol (A8511, Sigma), 2.5 mM p-Coumaric acid (C9008, Sigma) and 0.04% Hydrogen peroxide (w/v, L14000, Alpha Aesar, Tewksbury, MA) on a G:BOX (Syngene, Frederick, MD) equipped with a Synoptics 4.2MP camera and controlled with Genesys software (Syngene).

Figure: Abundance of PFKP in transformed H2009 cells upon CRISPR-based knockout (KO) of TRIM21. Representative data from two independent experiments. Please see Extended Data Figure 7e in the article [1].