IRF-3 antibody | knockout validation | Cell Signaling 4302

This is a knockout-validated antibody summary, based on the publication "Human Cytomegalovirus Immediate Early 86-kDa Protein Blocks Transcription and Induces Degradation of the Immature Interleukin-1β Protein during Virion-Mediated Activation of the AIM2 Inflammasome", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

IRF-3 antibody | knockout validation | Cell Signaling 4302 figure 1
Figure 1. Immunoblot assay for IRF3 and GAPDH in THP-1 cells transduced with Cas9 and gRNA targeting HUMCYC pseudogene or in indicated CRISPR-generated knockout cells. From [1].
Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: IRF-3

Catalog number: 4302

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxyl terminus of human IRF-3. Reacts with human, mouse, rat, and monkey. Suitable for western blot and immunoprecipitation.

Validation Method

Western blot

Sample

Control and IRF3/7-DKO THP-1 cells. Cell pellets were lysed in RIPA buffer (10 mM Tris-Cl (pH 8.0), 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, 0.1% SDS, 140 mM NaCl, and protease and phosphatase inhibitor cocktail).

Blocking agent

5% milk in 1× phosphate-buffered saline (PBS) containing 0.1% Tween 20 for 30 min at room temperature.

Primary incubation

Overnight at 4°C.

Secondary incubation

Horseradish peroxidase (HRP)-conjugated secondary antibodies in 5% milk for 30 min at room temperature.

Detection

Enhanced chemiluminescence (Thermo Fisher).

References
  1. Botto S, Abraham J, Mizuno N, Pryke K, Gall B, Landais I, et al. Human Cytomegalovirus Immediate Early 86-kDa Protein Blocks Transcription and Induces Degradation of the Immature Interleukin-1β Protein during Virion-Mediated Activation of the AIM2 Inflammasome. MBio. 2019;10: pubmed publisher