This is a knockout-validated antibody summary, based on the publication "Implementation of an antibody characterization procedure and application to the major ALS/FTD disease gene C9ORF72", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Cell lysates from HEK-293 parental, heterozygous (+/-) or two individual C9ORF72 KO clones (KOA, KOB) were prepared and processed for immunoblot with the indicated C9ORF72 antibody. The arrows point to positive C9ORF72 signals. The Ponceau stained transfers associated with each blot are shown as a protein loading control. lysis of 16 C9ORF72 antibodies by immunoblot.

Rabbit monoclonal IgG

Company: Abcam

Antibody: C9orf72

Catalog number: ab221137

Summary: Rabbit monoclonal IgG against a recombinant fragment within human C9orf72 (aa 1-250). Reacts with mouse, rat and human. Suitable for western blot. Unsuitable for flow cytometry, immunocytochemistry/immunofluorescence or immunohistochemistry (paraffin).

Figure 2. Longer exposure immunoblots on HEK-293 cell lysates and immunoblots of brain lysates from WT and C9ORF72 KO mice. The arrows point to positive C9ORF72 signals. From [1].

Western blot

HEK-293 parental, heterozygous (+/-) and C9ORF72 KO clones, and WT and C9ORF72 KO brain tissues. Cultured cells were collected in HEPES lysis buffer (20 mM HEPES, 100 mM sodium chloride, 1 mM EDTA, 5% glycerol, 1% Triton X-100, pH 7.4) supplemented with protease inhibitor.

5% milk in TBS with 0.1% Tween 20 (TBST).

O/N at 4˚C with 5% bovine serum albumin in TBS with 0.1% Tween 20 (TBST).

1:10,000 dilution anti-rabbit (Jackson ImmunoResearch Laboratories) in TBST with 5% milk for 1 hr at room temperature.

Antibody is not very specific for ICC/IF.

Detection of immuno-reactive bands was performed by image scan using a LI-COR Odyssey Imaging System (LI-COR Biosciences).