Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker

• WB

Lab

Western blot - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

Western blot : Anti-GJA1 antibody (ab11370) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab11370 was shown to bind specifically to GJA1. A band was observed at 45 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in GJA1 knockout cell line. To generate this image, wild-type and GJA1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (ab11370) at 1/2000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lanes 1 - 4:

Western blot - Human GJA1 knockout U-87 MG cell line (ab306676)

Lane 2:

GJA1 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type HEK-293 cell lysate at 20 µg

Lane 4:

GJA1 knockout HEK-293 <a href='/en-us/products/cell-lysates/human-gja1-knockout-hek-293-cell-lysate-ab261658'>ab261658</a> cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 45 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

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).

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

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. 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.

This image is courtesy of Carl Hobbs, King's College London, United Kingdom

• ICC

AbReview48192****

Immunocytochemistry - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

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).

This image is courtesy of an anonymous Abreview

• IHC-P

AbReview76879****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed human palatine tonsil. Stained with ab11370 at 1/1000 dilution. Secondary antibody used was Dako EnVision+ System- HRP Labelled Polymer Anti-R. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and 5% Normal Goat Serum for 19 hours at 4°C. Antigen retrieval method was heat mediated Tris/EDTA pH 9.0

This image is courtesy of an anonymous Abreview

• IHC-P

AbReview76880****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed rat heart (cardiac muscle tissue). Stained with ab11370 at 1/1000 dilution. Secondary antibody used was Dako EnVision+ System- HRP Labelled Polymer Anti-R. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and 5% Normal Goat Serum for 19 hours at 4°C. Antigen retrieval method was heat mediated Tris/EDTA pH 9.0

This image is courtesy of an anonymous Abreview

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

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.

This image is courtesy of Carl Hobbs, King's College London, United Kingdom

• IHC-P

AbReview76878****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed mouse spleen tissue. Stained with ab11370 at 1/1000 dilution. Secondary antibody used was Dako EnVision+ System- HRP Labelled Polymer Anti-R. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and 5% Normal Goat Serum for 19 hours at 4°C. Antigen retrieval method was heat mediated Tris/EDTA pH 9.0

This image is courtesy of an anonymous Abreview

• WB

Supplier Data

Western blot - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

All lanes:

Western blot - 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

Predicted band size: 43 kDa

false

• ICC

Supplier Data

Immunocytochemistry - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

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).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

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.

• WB

AbReview25925****

Western blot - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

All lanes:

Western blot - 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

Predicted band size: 43 kDa

Observed band size: 43 kDa

true

Exposure time: 30s

Image courtesy of an anonymous Abreview.

• WB

Supplier Data

Western blot - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (AB11370)

All lanes:

Western blot - Anti-Connexin 43 / GJA1 antibody - Intercellular Junction Marker (ab11370) at 1/8000 dilution

All lanes:

Mouse brain extract

Secondary

All lanes:

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

Predicted band size: 43 kDa

false