This is a knockout-validated antibody summary, based on the publication "Zinc-finger nuclease mediated disruption of Rag1 in the LEW/Ztm rat", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Western Blot analysis of the thymus from wild type, heterozygous and Rag1 mutant rats compared to GAPDH as housekeeping protein. From [1].

Antibody: RAG-1

Catalog number: sc-5599

Summary: Rabbit polyclonal IgG raised against the epitope corresponding to amino acids 744-1043 mapping at the C-terminus of RAG-1 of human origin. Recommended for detection of RAG-1 of mouse, rat and human origin by western blot, immunoprecipitation, immunofluorescence and ELISA; also reactive with additional species, including equine, canine, bovine, porcine and avian.

Company: SCBT

Western blot

Thymus from wild type, heterozygous and Rag1 mutant rats. NP40 buffer supplemented with Complete Mini Protease Inhibitor Cocktail Tablets (Roche, Mannheim, Germany), sodium orthovanadate (NEB, Frankfurt, Germany), PMSF (Sigma-Aldrich, Seelze, Germany) and DTT (Sigma-Aldrich, Seelze, Germany).

1 hour at room temperature with blocking solution (5% dry milk (Carl Roth, Karlsruhe, Germany) and 2.5% BSA, fraction V (Sigma-Aldrich, Seelze, Germany) in 3% TBS-Tween, pH 7,4).

1:300 dilution in blocking solution at 4C overnight.

1:2500 dilution of a donkey anti rabbit secondary antibody (ab16284, Abcam, Cambridge, UK) for 1 hour at room temperature in 3% TBS-Tween, pH 7.4.

Roti-Lumin was used as a chemoluminescence substrate.

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.