EFP antibody | knockout validation | BD Biosciences 610570

This is a knockout-validated antibody summary, based on publications "The Human Papillomavirus E6 Oncoprotein Targets USP15 and TRIM25 To Suppress RIG-I-Mediated Innate Immune Signaling" [1] (see figure 5b in the article), "Nuclear TRIM25 Specifically Targets Influenza Virus Ribonucleoproteins to Block the Onset of RNA Chain Elongation" [2] (see figure 2a in the article) and "TRIM25 Enhances the Antiviral Action of Zinc-Finger Antiviral Protein (ZAP)", as cited below [3]. All three articles indicate knockout-validation for Western blot. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

EFP antibody | knockout validation | BD Biosciences 610570 figure 1
Figure 1. Protein level of TRIM25 in the parental ZC3HAV1-knockout 293T, and CRISPR clones E, D and F. Clone E has similar TRIM25 expression as the parental cells whereas TRIM25 expression is significantly lower in clones D and F (designated TRIM25lo) that are mutated for wild type TRIM25. β-actin was used as a loading control. From [3].
Antibody information

Mouse monoclonal IgG1

Company: BD Biosciences

Antibody: EFP

Catalog number: 610570

Summary: Mouse monoclonal IgG1 against human EFP (TRIM25) aa. 32-210. Reacts with human by western blot, immunoprecipitation and immunofluorescence. Immunohistochemistry is not recommended.

Validation Method

Western blot

Sample

Control and TRIM25-KO ZC3HAV1-knockout 293T cells.

Primary incubation

1:5,000 dilution.

Secondary incubation

1:20,000 goat anti-mouse HRP (115-035-146; Jackson ImmunoResearch).

Detection

ECL Prime Western Blotting Detection Reagent (GE Healthcare) or SuperSignal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific).

References
  1. Chiang C, Pauli E, Biryukov J, Feister K, Meng M, White E, et al. The Human Papillomavirus E6 Oncoprotein Targets USP15 and TRIM25 To Suppress RIG-I-Mediated Innate Immune Signaling. J Virol. 2018;92: pubmed publisher
  2. Meyerson N, Zhou L, Guo Y, Zhao C, Tao Y, Krug R, et al. Nuclear TRIM25 Specifically Targets Influenza Virus Ribonucleoproteins to Block the Onset of RNA Chain Elongation. Cell Host Microbe. 2017;22:627-638.e7 pubmed publisher
  3. Li M, Lau Z, Cheung P, Aguilar E, Schneider W, Bozzacco L, et al. TRIM25 Enhances the Antiviral Action of Zinc-Finger Antiviral Protein (ZAP). PLoS Pathog. 2017;13:e1006145 pubmed publisher