Anti-beta Catenin antibody [15B8]

• WB

Project

Western blot - Anti-beta Catenin antibody [15B8] (AB6301)

This blot was produced using a 10% 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 5% Bovine Serum Albumin before being incubated with ab6301 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-beta Catenin antibody [15B8] (ab6301) at 1 µg/mL

Lane 1:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 3:

Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 5:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

Predicted band size: 85 kDa

Observed band size: 73 kDa,95 kDa

true

Exposure time: 4min

• WB

Lab

Western blot - Anti-beta Catenin antibody [15B8] (AB6301)

Lane 1 : HeLa whole cell lysate (20 µg) Lane 2 : NIH/3T3 whole cell lysate (20 µg) Lane 3 : PC12 whole cell lysate (20 µg) Lane 4 : Wild-type HAP1 whole cell lysate (20 µg) Lane 5 : Empty Lane Lane 6 : HPRT1 knockout HAP1 whole cell lysate (20 µg) Lanes 1 - 6 : Merged signal (red and green). Green - ab10479 observed at 24 kDa. Red - loading control ab6301 observed at 95 kDa. Gel type : MES Blocking buffer : LiCOR blocking buffer ab10479 was shown to specifically react with HPRT1 in wild-type HAP1 cells. No band was observed when HPRT1 knockout samples were examined. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE. ab10479 and ab6301 (Beta catenin - Loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800RD) preadsorbed and Goat anti-Mouse IgG H&L (IRDye® 680CW) preadsorbed secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HPRT antibody (<a href='/en-us/products/primary-antibodies/hprt-antibody-ab10479'>ab10479</a>) at 1 µg/mL

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

NIH/3T3 whole cell lysate at 20 µg

Lane 3:

PC12 whole cell lysate at 20 µg

Lane 4:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 6:

HPRT1 knockout HAP1 whole cell lysate at 20 µg

Secondary

Lanes 1 - 6:

Goat anti-Mouse IgG H&L (IRDye® 680CW) preadsorbed at 1/10000 dilution

Lanes 1 - 6:

Goat anti-Rabbit IgG H&L (IRDye® 800RD) preadsorbed at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-beta Catenin antibody [15B8] (AB6301)

Lanes 1 - 3 : Merged signal (red and green). Green - ab6301 observed at 85 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab6301 was shown to specifically react with beta-Catenin in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when knockout samples were used. Wild-type and beta Catenin knockout samples were subjected to SDS-PAGE. ab6301 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-beta Catenin antibody [15B8] (ab6301) at 1 µg/mL

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

CTNNB1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 85 kDa

false