This is a knockout-validated antibody summary, based on the publication "Comparative genetic screens in human cells reveal new regulatory mechanisms in WNT signaling", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

[enlarge]

Figure 1. Immunoblots of total AXIN1 and AXIN2 in various AXIN1 and/or AXIN2 deletion and/or truncation HAP1-7TGP cell lines. GAPDH loading controls are shown below their respective blots. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Axin2

Catalog number: 2151

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Pro566 of human Axin2. Reacts with human by western blot and immunoprecipitation.

Western blot

WT and AXIN2-KO HAP1-7TGP cells. Cell pellets were resuspended in 150 ml of ice-cold RIPA lysis buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 2% NP-40, 0.25% deoxycholate,

0.1% SDS, 1X SIGMAFAST protease inhibitors, 1 mM MgCl2, 62.5 U/ml Benzonase Nuclease, 1 mM DTT, 10% glycerol), and sonicated in a Bioruptor 300 (Diagenode) 4 x 30s in the high setting.

5% skim milk in TBST.

1:500 dilution in TBST + 5% skim milk overnight at 4˚C.

1:7,500 dilution peroxidase AffiniPure goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch Laboratories Cat. # 111-035-003) for 1 hr at room temperature.

SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific Cat. # 34095).