Anti-Claudin 1 antibody [EPR9306]

• WB

Lab

Western blot - Anti-Claudin 1 antibody [EPR9306] (AB180158)

Lanes 1 - 3 : Merged signal (red and green). Green - ab180158 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab180158 was shown to specifically react with in wild-type A431 WT cells as signal was lost in A431 CLDN1 knockout cells. Wild-type and A431 CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3pc Milk. ab180158 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Claudin 1 antibody [EPR9306] (ab180158) at 1/10000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CLDN1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CLDN1 knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-cldn1-knockout-a-431-cell-line-ab261889'>ab261889</a>)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 22 kDa

false

• WB

Supplier Data

Western blot - Anti-Claudin 1 antibody [EPR9306] (AB180158)

All lanes:

Western blot - Anti-Claudin 1 antibody [EPR9306] (ab180158) at 1/10000 dilution

Lane 1:

A431 cell lysate at 20 µg

Lane 2:

Human skin lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 22 kDa

false

• WB

Lab

Western blot - Anti-Claudin 1 antibody [EPR9306] (AB180158)

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

Claudin-1 expression is minimal in normal colon tissues and is increased in tumors (PMID : 17308096, PMID : 15965503).

All lanes:

Western blot - Anti-Claudin 1 antibody [EPR9306] (ab180158) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse colon tissue lysate at 20 µg

Lane 4:

Mouse small intestine tissue lysate at 20 µg

Lane 5:

NIH/3T3 (Mouse embryonic fibroblast) whole cell 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 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 18 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-Claudin 1 antibody [EPR9306] (AB180158)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

ab307692 may serve as a viable alternative to ab180158 in Western blot.

Lanes 1 - 6:

Western blot - Anti-Claudin 1 antibody [EPR25359-48] (<a href='/en-us/products/primary-antibodies/claudin-1-antibody-epr25359-48-ab307692'>ab307692</a>) at 1/1000 dilution

Lanes 7 - 12:

Western blot - Anti-Claudin 1 antibody [EPR9306] (ab180158) at 1/1000 dilution

Lanes 1 and 7:

Mouse kidney at 10 µg

Lanes 2 and 8:

Mouse liver at 10 µg

Lanes 3 and 9:

Mouse colon at 10 µg

Lanes 4 and 10:

Rat kidney at 10 µg

Lanes 5 and 11:

Rat liver at 10 µg

Lanes 6 and 12:

Rat colon 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 1/20000 dilution

Observed band size: 19 kDa

false

Exposure time: 60s

• WB

Supplier Data

Western blot - Anti-Claudin 1 antibody [EPR9306] (AB180158)

All lanes:

Western blot - Anti-Claudin 1 antibody [EPR9306] (ab180158) at 1/2000 dilution

All lanes:

Human fetal liver 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-ab136636'>ab136636</a>) at 1/500 dilution

Predicted band size: 22 kDa

false