This is a knockout-validated antibody summary, based on the publication "FMNL formins boost lamellipodial force generation", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. CRISPR/Cas9-mediated, specific loss of protein expression as documented by western blotting, using FMNL2/3-reactive antibody and tubulin as loading control. Asterisk marks non-specific band.

Mouse monoclonal IgG1kappa

Company: Abcam

Antibody: FMNL2

Catalog number: ab57963

Summary: Mouse monoclonal IgG1kappa against recombinant full length protein, corresponding to amino acids 1-179 of human FMNL2. Reacts with human by western blot.

Figure 2. Immunolabelling of FMNL2 and -3 confirms their absence in double-KO B16-F1 cell lines generated by CRISPR/Cas9. B16-F1 control as well as FMNL2/3 KO cells were stimulated with aluminum fluoride, immunolabelled for endogenous FMNL2 and -3 and counterstained for the actin cytoskeleton by fluorescent phalloidin. Bar is valid for all panels and equals 10 µm. From [1].

Western blot | Immunocytochemistry

B16 as well as FMNL2/3 KO cells. WB: cells were lysed in ice cold lysis-buffer (140 mM KCl, 50 mM Tris-HCl (pH 7.4), 50 mM NaF, 10 mM Na4P2O7, 2 mM MgCl2, 1% Triton-X100, supplemented with one mini complete protease inhibitor pill (Roche)). IC: cells were fixed with prewarmed, 4% paraformaldehyde (PFA) in PBS for 20 min

IC: 5% horse serum in 1% BSA in PBS.

WB: 1:1,000 dilution.

IC: 1:50 dilution.

WB: 1:10,000 dilution peroxidase-coupled anti-mouse IgG (Dianova; #115-035-062).

IC: Alexa Fluor 488- (1:400 dilution) or Alexa Fluor 594-coupled (1:200 dilution) anti-mouse IgG (Invitrogen; #A11029 or #A11032, respectively).