PKA-C- antibody | knockout validation | BD Biosciences 610981

This is a knockout-validated antibody summary, based on the publication "Local protein kinase A action proceeds through intact holoenzymes", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Mouse monoclonal IgG1

Company: BD Biosciences

Antibody: PKA-C

Catalog number: 610981

Summary: Mouse monoclonal IgG2b against human PKA-Cα subunit (aa. 18-347). Reacts with human, mouse, rat and dog. Suitable for western blot and immunofluorescence.

Validation Method

Western blot

Sample

WT and PKA R2A/R2B/CA triple knockout U2OS cells. Cells were harvested in lysis buffer (25 mM HEPES, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 20 mM NaF, 2% glycerol, 0.3% Triton X-100) containing 100 nM okadaic acid and protease inhibitors.

Primary incubation

1:1,000 dilution.

Secondary incubation

HRP-labeled secondary antibodies (Jackson Immunoresearch).

Detection

ECL (ThermoFisher) on an Alpha Innotech MultiImage III with FluoroChem Q software.

Figure

Immunoblots confirming knockout of PKA subunit proteins: (Top) RIIα, (top middle) RIIβ, ανδ (bottom middle) C subunit. (Bottom) Ponceau staining shows equal loading in WT and U2OSR2A/R2B/CA cells. Please see Figure 3b in the article [1].

References
  1. Smith F, Esseltine J, Nygren P, Veesler D, Byrne D, Vonderach M, et al. Local protein kinase A action proceeds through intact holoenzymes.. Science. 2017;356:1288-1293 pubmed publisher