This is a knockout-validated antibody summary, based on the publication "Dhh1 promotes autophagy-related protein translation during nitrogen starvation", 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. HEK293A WT or DDX6 KO cells were incubated in amino acid–free medium for the indicated times. Proteins were analyzed by immunoblotting. From [1].

Mouse monoclonal IgG2b (kappa light chain)

Company: Santa Cruz

Antibody: RCK

Catalog number: sc-376433

Summary: Mouse monoclonal IgG2b (kappa light chain) specific for an epitope mapping between amino acids 3-31 at the N-terminus of human RCK (DDX6). Recommended for detection of RCK of mouse, rat and human origin by western blot, immunoprecipitation, immunofluorescence and ELISA.

Western blot

WT and DDX6 KO HEK293A cells. Whole-cell lysates were prepared in cell lysis buffer (20 mM Tris pH 7.6, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 2.5 mM sodium phosphate, 1 mM glycerol 2-phosphate, 1 mM NaF, 1 mM NaVO4, 0.3% CHAPS, 1× EDTA-free protease inhibitor cocktail Roche, 05 892 791 001).

5% skim milk.

1:200 dilution.

1:2,000 dilution horseradish peroxidase–coupled secondary anti-mouse antibodies (1706516, Bio-Rad).

Chemiluminescence images were obtained using the LAS4000 (GE) system.