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).

Rabbit polyclonal

Company: Zymed

Antibody: Cx32

Catalog number: 34-5700

Summary: Rabbit polyclonal against a synthetic peptide derived from the C-terminal sequence of the human connexin 32 protein. Reacts with rat, mouse, and human. Suitable for western blot, and immunohistochemistry.

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: PBS; 10 mM phosphate, 154 mM NaCl).

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

1:125 dilution in blocking buffer overnight at 4°C.

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

Laboratories, PA; 1:5,000) diluted in blocking buffer for 1 to 3 h.

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

Please see Figure 3d in the article [1].

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.