This is a knockout-validated antibody summary, based on publications "Autochthonous tumors driven by Rb1 loss have an ongoing requirement for the RBP2 histone demethylase", as cited below [1], and "Histone demethylase KDM5A promotes tumorigenesis of osteosarcoma tumor" for western blot knockout validation (figure 5c) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: JARID1A

Catalog number: 3876

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to the human JARID1A protein. Reacts with human and mouse. It is predicted to react based on 100% sequence homology with rat and bovine. Suitable for western blot and immunoprecipitation.

Western blot

Heart from control and Rbp2-KO mice. Tissues were thawed very briefly on ice and homogenized using TissueLyser II (Qiagen) in EBC lysis buffer (50 mM Tris-Cl pH 8.0 containing 0.5% vol/vol Nonidet P-40 and 250 mM NaCl) supplemented with a protease inhibitor mixture (Roche).

5% nonfat milk in TBST (20 mM Tris-Cl pH 7.6 containing 150 mM NaCl and 0.1% Tween-20) for 1 h at room temperature.

1:1,000 dilution in 5% BSA in TBST overnight at 4 °C.

At room temperature for 1 h.

Immobilon Western Chemiluminescent HRP Substrate (EMD Millipore) and developed using an ImageQuant LAS4000 chemiluminescence detector (GE Healthcare Life Sciences).

Immunoblot analysis of RBP2 expression in heart tissues harvested from Rbp2lox/lox and Rbp2lox/lox; Cre–ER mice that were treated as in C and D. Please see Suppl. Figure 1e in the article [1].