HRP Anti-5T4 antibody [EPR5530]

• WB

Lab

Western blot - HRP Anti-5T4 antibody [EPR5530] (AB199547)

ab199547 was shown to react with 5T4 (HRP) in wild-type MCF7 cells in western blot. Loss of signal was observed when TPBG knockout cell line ab269499 (knockout cell lysate ab269661) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab199547 overnight at 4 °C at a 1 in 5000 dilution Blots were developed with Optiblot ECL reagent (ab133456) and imaged.

All lanes:

Western blot - HRP Anti-5T4 antibody [EPR5530] (ab199547) at 1/5000 dilution

Lane 1:

Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TPBG knockout MCF7 cell lysate (<a href='/en-us/products/cell-lysates/human-tpbg-knockout-mcf7-cell-lysate-ab269661'>ab269661</a>) at 20 µg

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 85 kDa

false

• WB

Lab

Western blot - HRP Anti-5T4 antibody [EPR5530] (AB199547)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab199547 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-5T4 antibody [EPR5530] (ab199547) at 1/5000 dilution

All lanes:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Predicted band size: 46 kDa

Observed band size: 75 kDa

true

Exposure time: 20min