Anti-MUC4 antibody [EPR27199-56]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC4 antibody [EPR27199-56] (AB307546)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscl 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] (AB307546)

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] (AB307546)

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] (AB307546)

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] (ab307546) 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] (AB307546)

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] (ab307546) 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] (AB307546)

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] (ab307546) 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