This is a knockout-validated antibody summary, based on the publication "Genome-wide CRISPR screening reveals genetic modifiers of mutant EGFR dependence in human NSCLC", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Immunoblots of indicated proteins in control or RHOA knockout HCC827 cells. β-Actin was used as a loading control. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: RhoA

Catalog number: 2117

Summary: Rabbit monoclonal IgG against a synthetic peptides corresponding to residues near the carboxy terminus of human RhoA. Reacts with human, mouse, rat, monkey, and bovine. Suitable for western blot.

Western blot

Control or RHOA knockout HCC827 cells. Cells were lysed in RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) (Thermo Fisher Scientific #89901) supplemented with 100 x Protease Inhibitor Cocktail (Sigma #P8340), 100 x Phosphatase Inhibitor Cocktail (Thermo Fisher Scientific #78427) and 25 units/ml Benzonase Nuclease (Sigma #E8263).

5% Blotting-Grade Blocker (Bio-Rad #1706404) in Tris Buffered Saline with Tween 20, pH 8.0 (TBST) (Sigma #T9039) for 1 hr at room temperature.

1:1,000 dilution in 5% BSA (Akron Biotechnology #AK8917-0100) overnight at 4°C.

Horseradish peroxidase (HRP) conjugated secondary antibody diluted in 5% Blotting-Grade Blocker.

Amersham ECL Western Blotting Detection Reagents (GE Healthcare #RPN2106) or SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific #34580) and Amersham Hyperfilm ECL (GE Healthcare #28906839).