Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free

• Flow Cyt

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Flow Cytometry - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

Flow cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labelling MUC1 with unpurified ab45167 (pink) at a dilution of 1/150. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (green).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

• Flow Cyt

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Flow Cytometry - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

Flow Cytometry analysis of A549 (human lung carcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/20 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A goat anti-rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000 dilution. Black - Isotype control, Rabbit monoclonal IgG. Blue - unlabeled control, cells without incubation with primary antibody and secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

• IP

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Immunoprecipitation - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

ab45167 (purified) at 1/20 dilution (2ug) immunoprecipitating MUC1 in Human fetal lung lysate.

Lane 1 (input) : Human fetal lung lysate 10ug

Lane 2 (+) : ab45167 + Human fetal lung lysate 10ug

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab45167 in Human fetal lung lysate

For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection at 1/1000 dilution.

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

All lanes:

Immunoprecipitation - Anti-MUC1 antibody [EP1024Y] (<a href='/en-us/products/primary-antibodies/muc1-antibody-ep1024y-ab45167'>ab45167</a>)

Predicted band size: 122 kDa

Observed band size: 27 kDa

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• WB

Lab

Western blot - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MUC1 antibody [EP1024Y] (<a href='/en-us/products/primary-antibodies/muc1-antibody-ep1024y-ab45167'>ab45167</a>) at 1/2000 dilution

All lanes:

T47D cell lysate at 10 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 1/1000 dilution

Predicted band size: 122 kDa

Observed band size: 27 kDa

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Exposure time: 3min

• WB

Lab

Western blot - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

This data was developed using the same antibody clone in a different buffer formulation (ab45167).

Lanes 1- 2 : Merged signal (red and green). Green - ab45167 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab45167 was shown to react with MUC1 in Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cells in western blot. Loss of signal was observed when knockout cell line ab255412 (knockout cell lysate ab263764) was used. Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) and MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab45167 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MUC1 antibody [EP1024Y] (<a href='/en-us/products/primary-antibodies/muc1-antibody-ep1024y-ab45167'>ab45167</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 2:

Western blot - Human MUC1 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-muc1-knockout-hela-cell-lysate-ab263764'>ab263764</a>) at 20 µg

Predicted band size: 122 kDa

Observed band size: 24 kDa

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• WB

Lab

Western blot - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

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

All lanes:

Western blot - Anti-MUC1 antibody [EP1024Y] (<a href='/en-us/products/primary-antibodies/muc1-antibody-ep1024y-ab45167'>ab45167</a>) at 1/1000 dilution

Lane 1:

Human colon cancer lysate at 20 µg

Lane 2:

Rat liver lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 27 kDa

false

• WB

Lab

Western blot - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (AB218998)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

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

All lanes:

Western blot - Anti-MUC1 antibody [EP1024Y] (<a href='/en-us/products/primary-antibodies/muc1-antibody-ep1024y-ab45167'>ab45167</a>) at 1/5000 dilution

All lanes:

Mouse liver lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 27 kDa

false