Immunohistochemical analysis of formalin-fixed, paraffion-embedded cow heart tissue staining Connexin 43 / GJA1 using ab11370 at1/2000 dilution. Detected using an Anti-Rabbit IgG-biotin antibody and an Avidin-peroxidase conjugate.

All lanes : Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (ab11370) at 1/2000 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
Lane 2 : P19 (mouse embryonic carcinima cell line) cell lysate
Lane 3 : Rat2 (rat embryo-derived cell line) cell lysate
Lane 4 : MDCK (canine kidney cell line) cell lysate

Secondary
All lanes : Goat Anti-Rabbit IgG-peroxidase conjugate

Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (ab11370) at 1/8000 dilution + Mouse brain extract

Secondary
Anti-Rabbit IgG-AP conjugate with NBT/BCIP substrate

Immunocytochemical analysis of BHK (baby hamster kidney cell line) cells fixed and permeabilized with methanol followed by methanol:acetone. -Connexin 43 / GJA1 was labeled using ab11370 at 1/400 dilution, followed by a Goat Anti-Rabbit-FITC conjugate (Green). The nuclear counterstain is DAPI (Blue).

Immunohistochemical analysis of formalin-fixed, paraffion-embedded human heart tissue staining Connexin 43 / GJA1 using ab11370 at 1/2000 dilution, followed by a Goat Anti-Rabbit IgG-FITC conjugate (Green).

Immunohistochemical detection of Connexin 43 / GJA1 using ab11370 (1/1000) on human testis sections (PFA-fixed paraffin-embedded sections). Antigen retrieval : Heat mediated - Buffer/Enzyme Used: Citric acid pH6, 1% BSA as blocking agent for 10 mins @ rt°C. Biotin labelled secondary antibody used at 1/200. Stained are clusters of Leydig cells in the interstitium and what should be Sertoli cells within the seminiferous tubules and the junction between (?). Sertoli cells and tubule "capsular" cells are intensely positive at their membrane interfaces (capsular interface positivity only evident in upper middle of image). Interestingly,smooth muscle of human colon and myometrium were completely negative when tested at the same time.

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All lanes : Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (ab11370) at 1/6000 dilution

Lane 1 : Whole tissue lysate prepared from murine left ventricle at 5 µg
Lane 2 : Whole tissue lysate prepared from murine left ventricle at 10 µg
Lane 3 : Whole tissue lysate prepared from murine left ventricle at 20 µg
Lane 4 : Whole tissue lysate prepared from murine left ventricle at 30 µg
Lane 5 : Whole tissue lysate prepared from murine left ventricle at 40 µg

Secondary
All lanes : HRP-conjugated donkey anti-rabbit polyclonal at 1/10000 dilution

Developed using the ECL technique.

Observed band size: 43 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

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ab11370 staining Connexin 43 / GJA1 in human pluripotent stem cell derived cardiomyocyte by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with saponin and blocked with 5% serum for 15 minutes at room temperature. Samples were incubated with primary antibody (1/1000) for 16 hours at 4°C. An Alexa Fluor^® 568-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

Clear antibody signal (yellow) at the interface of adjoining cells in a population of human embryonic stem cell derived cardiomyocytes (red = sarcomeric alpha actinin).

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Immunohistochemistical detection of  Connexin 43 / GJA1 using ab11370 on Formaldehyde-fixed paraffin-embedded mouse heart muscle sections. Antigen retrieval step: heat mediated using citric acid pH6. Blocking step: 1% BSA for 10 mins @ rt°C.  Primary antibody ab11370 incubated at 1/2000 for 2 hours.  Secondary antibody: anti rabbit IgG conjugated to biotin used at 1/200. Submitted image of cardiac myofibres in L/S shows clear, specific labelling of Intercalated discs which are rich in Gap junctions (Connexin 43 is a major component of Gap junctions). No other positivity is observed in mouse heart (the image is taken from a whole heart section). NB: in mouse tongue the ventral keratinising stratified squamous epithelium shows positivity in not only basal cells but also in the Prickle cell layer, although markedly reduced: data not shown but see here http://www.immunoportal.com/ for appropriate image.