This is a knockout-validated antibody summary, based on the publication "Importance of Validating Antibodies and Small Compound Inhibitors Using Genetic Knockout Studies-T Cell Receptor-Induced CYLD Phosphorylation by IKKε/TBK1 as a Case Study", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Wild-type (wt) or three different CYLD knockout (ko) Jurkat T cell clones were stimulated with 200 ng/ml PMA and 1µM Ionomycin for the indicated times. From [1].

Mouse monoclonal IgG2a (kappa light chain)

Company: Santa Cruz

Antibody: cylindromatosis 1

Catalog number: sc-74435

Summary: Mouse monoclonal IgG2a (kappa light chain) against the C-terminal 419 amino acids of human cylindromatosis 1. Recommended for detection of cylindromatosis 1 of mouse, rat and human origin by western blot, immunoprecipitation, immunofluorescence and ELISA.

Western blot

Wild-type and CYLD knockout Jurkat T cell clones.

5% milk powder in TBS/0.2% Tween 20 (TBST) for 1 h at room temperature (RT).

1:500 dilution overnight at 4◦C in 5% milk powder in TBST.

HRP-conjugated secondary antibodies (GE healthcare) in 5% milk powder in TBST for 1h at RT.

Western Lightning ECL detection system (Perkin Elmer Life Sciences) according to the manufacturer’s instructions.