This is a knockout-validated antibody summary, based on the publication "Testing for a gap junction-mediated bystander effect in retinitis pigmentosa: secondary cone death is not altered by deletion of connexin36 from cones", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Cx36 antibody staining in vertical sections of wild-type (wt) mice (B), Rho−/− (pw5) (C) and rd1 mice (p21) (E). The staining is absent in Rho−/− and rd1 mice with a targeted deletion of Cx36 (D, F). Cx36 immunoreactivity in magnified regions in the outer plexiform layer (OPL) of wt (H), Rho−/−Cx36+/+ (I) and rd1Cx36+/+ mice (K) produced fine punctuate labeling in the outer plexiform layer (OPL) where rods and cones are electrically coupled by Cx36. This staining was absent in the OPL of Cx36-deficient Rho−/− (J) and rd1 mutants (L). Residual staining (arrows) was caused by unspecific binding of the secondary antibody to blood vessels and was also present in controls (stainings without primary antibody). Retinal layers are indicated in the Nomarski micrographs (A, G). Scale bars = 10 µm in F (applies to A–F); in L (applies to G–L). From [1].

Rabbit polyclonal IgG

Company: Invitrogen

Antibody: Connexin36

Catalog number: 51-6300

Summary: Rabbit polyclonal IgG against a peptide corresponding to a sequence from the C-terminus of rat and mouse Connexin 36 (Cx36). Reacts with human, mouse, and rat. Suitable for western blot, immunoprecipitation and ELISA, as well as immunohistochemistry (IHC) and immunocytochemistry (ICC).

Immunohistochemistry

Vertical sections of wild-type, Rho−/−Cx36+/+, Rho−/−Cx36-/-, rd1Cx36+/+ , and rd1Cx36-/- mice. Eyeballs were fixed in 2% PFA (PFA; Riedel de Haen, Seelze, Germany) or for 60 minutes in 4% PFA in phosphate-buffered saline, pH 7.4 (PBS), embedded in cryoblock (Medite GmbH, Burgdorf, Germany) at −20°C. Vertical sections (18–20 µm for morphological analysis; 12 µm for quantitative analysis) were cut on a cryostat (Bright, Huntingdon, United Kingdom).

10% normal goat serum (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) or 10% normal donkey serum (Dianova GmbH, Hamburg, Germany) in TBST.

1∶500 dilution overnight at 4°C.

Alexa Fluor 488 or 568 (Invitrogen, Karlsruhe, Germany) or Cy3 (Jackson Immunoresearch, West Grove, PA) diluted in blocking solution for at least two hours.

Leica TCS SL confocal microscope with a 40×/1.25 oil plan apochromat or with a 63×/1.32 oil plan apochromat objective (z-axis step size 0.2 µm).

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.