This is a knockout-validated antibody summary, based on the publication "A Genome-wide Haploid Genetic Screen Identifies Regulators of Glutathione Abundance and Ferroptosis Sensitivity", as cited below [1], and "KEAP1 deficiency drives glucose dependency and sensitizes lung cancer cells and tumors to GLUT inhibition" for western blot knockout validation (figure 1c) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Kelch-like ECH associated protein 1 (KEAP1), MRP1, and NRF2 protein levels in HAP1 Control and gene-disrupted cells. From [1].

Mouse monoclonal IgG2b (kappa light chain)

Company: Santa Cruz

Antibody: Keap1

Catalog number: sc-365626

Summary: Mouse monoclonal IgG2b (kappa light chain) against amino acids 181-370 mapping within an internal region of human Keap1. Recommended for detection of human Keap1 by western blot, immunoprecipitation, immunofluorescence and ELISA.

Western blot

Control and KEAP1-KO HAP1 cells. Cells were lysed in a modified RIPA buffer (50 mM Tris pH 7.5, 150mM NaCl, 0.5% NP-40, 0.1% Sodium deoxycholate, 1% Triton X-100, 0.1% SDS, 2 mM EDTA) with complete protease inhibitor cocktail (Cat# P8340, Sigma) on ice for 20 min.

1:500 dilution in Odyssey Buffer (Cat# 927-40100, LI-COR Biotechnology, Lincoln, NE) at 60 rpm, 4°C, overnight.

1:15,000 dilution donkey anti-mouse-680 or donkey anti-mouse-800 (Cat# 926-68022 or Cat# 926-32212, LI-COR) in Odyssey Buffer for 40 min, 60 rpm, RT.

LI-COR Odyssey CLx imager.