Cyclin D1 antibody | knockout validation | Cell Signaling 2978

This is a knockout-validated antibody summary, based on the publication "Cyclin D-CDK4 kinase destabilizes PD-L1 via cullin 3-SPOP to control cancer immune surveillance", 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: Cyclin D1

Catalog number: 2978

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to the carboxy-terminus of human cyclin D1. Reacts with human, mouse and rat. Suitable for western blot, immunohistochemistry (paraffin) and flow cytometry.

Validation Method

Western blot

Sample

Wild type and combinational (cyclin D1−/−D2−/−D3−/−) or single isoform cyclin D knockout MEFs. Cells were lysed in EBC buffer (50 mM Tris pH 7.5, 120 mM NaCl, 0.5% NP-40) supplemented with protease inhibitors (Complete Mini, Roche) and phosphatase inhibitors (phosphatase inhibitor cocktail set I and II, Calbiochem).

Secondary incubation

Peroxidase-conjugated anti-rabbit secondary antibody (Sigma, A-4914).

Figure

Immunoblot analysis of whole cell lysates derived from wild type versus combinational (cyclin D1−/−D2−/−D3−/−) (a) or single isoform cyclin D knockout MEFs. Please see Figure 2a,b in the article [1].

References
  1. Zhang J, Bu X, Wang H, Zhu Y, Geng Y, Nihira N, et al. Cyclin D-CDK4 kinase destabilizes PD-L1 via cullin 3-SPOP to control cancer immune surveillance. Nature. 2018;553:91-95 pubmed publisher