Anti-Cx40/GJA5 antibody [EPR28370-62]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Cx40/GJA5 with ab313644 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse pancreas.The section was incubated with ab313644 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 LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse colon (fresh) tissue labeling Cx40/GJA5 with ab313644 at 1/50 (10.32 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on mouse colon. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313644 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse pancreas (fresh) tissue labeling Cx40/GJA5 with ab313644 at 1/50 (10.32 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on mouse pancreas. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313644 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Cx40/GJA5 with ab313644 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on endothelium of mouse lung (PMID : 17483738).The section was incubated with ab313644 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 LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of paraffin-embedded Mouse atrium cardiac tissue labeling Cx40/GJA5 with ab313644 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse atrium cardiac muscle (PMID : 30564139, 17122196).The section was incubated with ab313644 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 LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Cx40/GJA5 with ab313644 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse kidney juxtaglomerulus (PMID : 29427253).The section was incubated with ab313644 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 LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh) tissue labeling Cx40/GJA5 with ab313644 at 1/50 (10.32 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on endothelium of mouse lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313644 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IP

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Immunoprecipitation - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

Cx40/GJA5 was immunoprecipitated from 0.35 mg Mouse lung tissue lysate with ab313644 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313644 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse lung tissue lysate Lane 2 : ab313644 IP in Mouse lung tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313644 in mouse lung tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds

All lanes:

Immunoprecipitation - Anti-Cx40/GJA5 antibody [EPR28370-62] (ab313644) at 1/30 dilution

All lanes:

Mouse lung tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

false

Exposure time: 24s

• WB

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Western blot - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

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

Negative control : colon, stomach.

The molecular weight observed is consistent with what has been described in the literature (PMID : 29285063, 19726871).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

Exposure time : Lane 1-3 : 37 seconds; Lane 4 : 10 seconds.

All lanes:

Western blot - Anti-Cx40/GJA5 antibody [EPR28370-62] (ab313644) at 1/1000 dilution

Lane 1:

Mouse heart tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse stomach tissue lysate at 20 µg

Lane 4:

Mouse lung tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 40 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Cx40/GJA5 antibody [EPR28370-62] (AB313644)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 29285063, 19726871).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Anti-Integrin alpha V (ab179475) control staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Cx40/GJA5 antibody [EPR28370-62] (ab313644) at 1/1000 dilution

Lane 1:

Mouse lung non-membrane fraction at 20 µg

Lane 2:

Mouse lung membrane fraction at 20 µg

Secondary

All lanes:

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

Observed band size: 40 kDa,36 kDa

false

Exposure time: 37s