This is a knockout-validated antibody summary, based on the publication "Tissue-specific cross-reactivity of connexin32 antibodies: problems and solutions unique to the central nervous system", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG1, kappa

Company: Zymed

Antibody: Cx32

Catalog number: 13-8200

Summary: Mouse monoclonal IgG1 against a synthetic peptide corresponding to a segment of the cytoplasmic loop of rat Connexin-32. It reacts with human, mouse, rat, and sheep. Recommended for western blot, immunofluorescence, immunohistochemistry (frozen) and ELISA.

Western blot

Brain and liver tissues from human and WT, and Cx32 knockout mice. All tissues were homogenized in fresh RIPA buffer (1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM NaF, 50 g/ml aprotinin, 1 mM sodium orthovanadate, 1 mg/ml phenylmethylsulfonyl fluoride, 10 mM phosphate-buffered saline [2] ).

5% (w/v) skim milk powder–PBS with 0.1% Tween-20 (PBST; blocking buffer) for 1 to 3 h.

1:250 dilution in blocking buffer overnight at 4°C.

Horseradish peroxidase (HRP)-conjugated anti-mouse (Jackson ImmunoResearch

Laboratories, PA; 1:2,000) diluted in blocking buffer for 1 to 3 h.

X-ray film using SuperSignal West Pico Chemiluminescent Substrate (Pierce, IL).

Please see Figure 2d in the article [1].

This antibody is specific in liver tissue but cross-reacts with a protein(s) in both Cx32+/+ and Cx32−/− control brain tissue. Note that this particular cross-reactive protein(s) migrates with the same mobility as Cx32 when separated on 12.5% Tris-HCl polyacrylamide gels.

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.