重组Anti-Connexin 43 / GJA1抗体[EPR22955-101] - Intercellular Junction Marker

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling Connexin 43 / GJA1 with ab235585 at 1/500 dilution (1.01 μg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Positive staining on human myocardial intercalated disc is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse cardiac muscle tissue staining Connexin 43 / GJA1 with ab235585 at 0.507 μg/ml, ab308619 anti-MYBPC3, and ab218198 anti-Dystrophin.

Panel A : anti-Connexin 43 / GJA1 (green; Opal™520), anti-MYBPC3 (magenta; Opal™570), anti-Dystrophin (gray; Opal™690) on mouse cardiac muscle.

Panel B : anti-Connexin 43 / GJA1 staining intercalated discs in mouse cardiac muscle.

Panel C : anti-MYBPC3 staining cytoplasm of heart myocytes in mouse cardiac muscle.

Panel D : anti-Dystrophin staining membrane of heart myocytes in mouse cardiac muscle.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab235585, ab308619 and ab218198 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat cardiac muscle tissue staining Connexin 43 / GJA1 with ab235585 at 0.507 μg/ml, ab308619 anti-MYBPC3, and ab218198 anti-Dystrophin.

Panel A : anti-Connexin 43 / GJA1 (green; Opal™520), anti-MYBPC3 (magenta; Opal™570), anti-Dystrophin (gray; Opal™690) on rat cardiac muscle.

Panel B : anti-Connexin 43 / GJA1 staining intercalated discs in rat cardiac muscle.

Panel C : anti-MYBPC3 staining cytoplasm of heart myocytes in rat cardiac muscle.

Panel D : anti-Dystrophin staining membrane of heart myocytes in rat cardiac muscle.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab235585, ab308619 and ab218198 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse cardiac muscle tissue staining Connexin 43 / GJA1 with ab235585 at 0.507 μg/ml, ab308619 anti-MYBPC3, and ab259345 anti-Laminin subunit alpha-2.

Panel A : anti-Connexin 43 / GJA1 (green; Opal™520), anti-MYBPC3 (magenta; Opal™570), anti-Laminin subunit alpha-2 (gray; Opal™690) on mouse cardiac muscle.

Panel B : anti-Connexin 43 / GJA1 staining intercalated discs in mouse cardiac muscle.

Panel C : anti-MYBPC3 staining cytoplasm of heart myocytes in mouse cardiac muscle.

Panel D : anti-Laminin subunit alpha-2 staining basal laminae in mouse cardiac muscle.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab235585, ab308619 and ab259345 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labeling Connexin 43 / GJA1 with ab235585 at 1/1000 dilution (0.507 μg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Positive staining on rat myocardial intercalated disc is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labeling Connexin 43 / GJA1 with ab235585 at 1/1000 dilution (0.507 μg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Positive staining on mouse myocardial intercalated disc (PMID : 30228305) is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of mouse cardiac muscle tissue labeling Connexin 43 / GJA1 using ab235585 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). The nuclear counterstain is DAPI (blue). Positive staining on intercalated discs of mouse cardiac muscle (PMID : 18678643) is observed.

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Connexin 43 / GJA1 with ab235585 at 1/1000 dilution (0.507 μg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Membranous staining on mouse testis is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of rat cardiac muscle tissue labeling Connexin 43 / GJA1 using ab235585 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). The nuclear counterstain is DAPI (blue). Positive staining on intercalated discs of rat cardiac muscle (PMID : 18678643) is observed.

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IP

Unknown

Immunoprecipitation - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Connexin 43 / GJA1 was immunoprecipitated from 0.35 mg mouse brain lysate 10μg with ab235585 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab235585 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain lysate 10μg.

Lane 2 : ab235585 IP in mouse brain lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab235585 in mouse brain lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

High MW band between 75kDa and 100kDa represents Connexin 43 dimers which described in literature. (PMID : 19460776).

All lanes:

Immunoprecipitation - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (ab235585)

Predicted band size: 43 kDa

Observed band size: 43 kDa

false

• WB

Lab

Western blot - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Blocking and Dilution Buffer and concentration : 5% NFDM/TBST. The expression profile is consistent with the existence of natural variants described in the literature (PMID : 24210816, 28576298). Exposure times : Lane 1 : 59 seconds; Lane 2 : 15 seconds; Lane 3 : 15 seconds; Lane 4 : 26 seconds.

All lanes:

Western blot - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (ab235585) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 10 µg

Lane 4:

Human brain tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 43 kDa

Observed band size: 43 kDa

false

• WB

Lab

Western blot - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

ab235585 has low affinity in cell line samples.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (ab235585) at 1/1000 dilution

Lane 1:

Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Predicted band size: 43 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Connexin 43 / GJA1 antibody [EPR22955-101] - Intercellular Junction Marker (AB235585)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18649180; PMID : 30448479). Alkaline phosphatase. - : untreated membrane; + : phosphatase treated membrane. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab235585) total protein control staining at 1/1000 dilution. Exposure time : 70 seconds

All lanes:

Western blot - Anti-Connexin 43 / GJA1 (phospho Y265) antibody [EPR26416-134] (<a href='/products/primary-antibodies/connexin-43-gja1-phospho-y265-antibody-epr26416-134-ab312836'>ab312836</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) cells transfected with an empty vector containi a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

HeLa cells transfected with a RSV-SR-A V-SRC expression vector containi a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

HeLa cells transfected with a mouse GJA1 expression vector containi a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

HeLa cells co-transfected with a mouse GJA1 expression vector containi a myc-His-tag® and a RSV-SR-A V-SRC expression vector containi a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 48 kDa,55 kDa

false

Exposure time: 70s