Rad23B antibody | knockout validation | Cell Signaling 13525

This is a knockout-validated antibody summary, based on the publication "Stress- And Ubiquitylation-Dependent Phase Separation of the Proteasome", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Rad23B

Catalog number: 13525

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Ala140 of human Rad23B protein. Reacts with human and monkey. Suitable for western blot.

Validation Method

Western blot

Sample

Wild-type and RAD23B-KO HCT116 cells. Cells were lysed with buffer A with complete protease inhibitor cocktail (EDTA free) (Roche) for 30 min on ice.

Blocking agent

5% non-fat milk for 30 min.

Primary incubation

1 h at room temperature.

Secondary incubation

HRPconjugated goat anti-rabbit Ig (Jackson ImmunoResearch Laboratories) for 30 min at room temperature.

Detection

ECL Prime Western Blotting Detection Reagent (GE Healthcare) and analysed on an ImageQuant LAS4000 (GE Healthcare).

Figure

Wild-type, RAD23B-KO (RAD23BKO/KO) or UBE3A-KO (UBE3AKO/KO) HCT116 cells were stimulated with 0.2 M sucrose for 30 min and immunoblotted with the indicated antibodies. Please see Extended Data Figure 8a in the article [1].

References
  1. Yasuda S, Tsuchiya H, Kaiho A, Guo Q, Ikeuchi K, Endo A, et al. Stress- and ubiquitylation-dependent phase separation of the proteasome. Nature. 2020;578:296-300 pubmed publisher