Anti-MUC4 antibody [EPR27199-56] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC4 antibody [EPR27199-56] - BSA and Azide free (AB307547)

This data was developed using ab307546, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling MUC4 with ab307546 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on human skeletal muscle. The section was incubated with ab307546 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) was used. 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-MUC4 antibody [EPR27199-56] - BSA and Azide free (AB307547)

This data was developed using ab307546, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MUC4 with ab307546 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human colon. The section was incubated with ab307546 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) was used. 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-MUC4 antibody [EPR27199-56] - BSA and Azide free (AB307547)

This data was developed using ab307546, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cervical carci tissue labeling MUC4 with ab307546 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human cervical carcinoma. The section was incubated with ab307546 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-MUC4 antibody [EPR27199-56] - BSA and Azide free (AB307547)

This data was developed using ab307546, the same antibody clone in a different buffer formulation.

MUC4 was immunoprecipitated from 0.35 mg Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab307546 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307546 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab307546 IP in Capan-1 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307546 in Capan-1 whole cell lysate.

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

Exposure time : 93 seconds.

All lanes:

Immunoprecipitation - Anti-MUC4 antibody [EPR27199-56] (<a href='/en-us/products/primary-antibodies/muc4-antibody-epr27199-56-ab307546'>ab307546</a>) at 1/1000 dilution

Lane 1:

Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2:

Capan-1 whole cell 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 1/5000 dilution

Predicted band size: 80 kDa

Observed band size: 100 kDa,170 kDa

false

Exposure time: 93s

• WB

Supplier Data

Western blot - Anti-MUC4 antibody [EPR27199-56] - BSA and Azide free (AB307547)

This data was developed using 307546, the same antibody clone in a different buffer formulation.

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

Negative control : skeletal muscle.

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.

The identity of the bands below 75 kDa are unknown.

The detected signal is the beta chain of MUC4.

All lanes:

Western blot - Anti-MUC4 antibody [EPR27199-56] (<a href='/en-us/products/primary-antibodies/muc4-antibody-epr27199-56-ab307546'>ab307546</a>) at 1/1000 dilution

Lane 1:

Human stomach tissue lysate 20 μg

Lane 2:

Human skeletal muscle cytoplasma tissue lysate 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

Predicted band size: 80 kDa

Observed band size: 100 kDa,170 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MUC4 antibody [EPR27199-56] - BSA and Azide free (AB307547)

This data was developed using 307546, the same antibody clone in a different buffer formulation.

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

Negative control : MIA PaCa-2 (PMID : 21886786), PANC-1 (PMID : 23152882)

The detected signal is the beta chain of MUC4.

All lanes:

Western blot - Anti-MUC4 antibody [EPR27199-56] (<a href='/en-us/products/primary-antibodies/muc4-antibody-epr27199-56-ab307546'>ab307546</a>) at 1/1000 dilution

Lane 1:

Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 20 &mu;g

Lane 2:

MIA PaCa-2 (human pancreas epithelial cell) whole cell lysate 20 &mu;g

Lane 3:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate 20 &mu;g

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

Predicted band size: 80 kDa

Observed band size: 100 kDa,170 kDa

false

Exposure time: 147s