Anti-beta Dystroglycan antibody [EP2200Y] - BSA and Azide free

• WB

Lab

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] - BSA and Azide free (AB284707)

This data was developed using ab62373, the same antibody clone in a different buffer formulation.

Lanes 1-3 : Merged signal (red and green). Green - ab62373 observed at 43 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab62373 Anti-beta Dystroglycan antibody [EP2200Y] was shown to specifically react with beta Dystroglycan in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266263 (knockout cell lysate ab257192) was used. Wild-type and beta Dystroglycan knockout samples were subjected to SDS-PAGE. ab62373 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] (<a href='/en-us/products/primary-antibodies/beta-dystroglycan-antibody-ep2200y-ab62373'>ab62373</a>) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human DAG1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-dag1-knockout-hek-293t-cell-line-ab266263'>ab266263</a>)

Lane 2:

DAG1 knockout HEK293T cell lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Predicted band size: 97 kDa

Observed band size: 43 kDa

false

• WB

Lab

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] - BSA and Azide free (AB284707)

This data was developed using the same antibody clone in a different buffer formulation (ab62373)

ab62373 was shown to react with DAG1 in wild-type A431 cells in Western blot with loss of signal observed in DAG1 knockout cell line ab261906. Wild-type A431 and DAG1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab62373 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] (<a href='/en-us/products/primary-antibodies/beta-dystroglycan-antibody-ep2200y-ab62373'>ab62373</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 lysate at 20 µg

Lane 2:

Western blot - Human DAG1 knockout A-431 cell line (ab261906) at 20 µg

Observed band size: 43 kDa

false

• WB

Lab

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] - BSA and Azide free (AB284707)

This data was developed using ab62373, the same antibody clone in a different buffer formulation.

Blocking Buffer : 5% NFDM /TBST

All lanes:

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] (<a href='/en-us/products/primary-antibodies/beta-dystroglycan-antibody-ep2200y-ab62373'>ab62373</a>) at 0.526 µg/mL

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Mouse heart lysate at 20 µg

Lane 3:

Mouse skeletal muscle at 20 µg

Lane 4:

Neuro-2a (Mouse neuroblastoma cell line) whole cell 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 0.05 µg/mL

Predicted band size: 97 kDa

false

• WB

Unknown

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] - BSA and Azide free (AB284707)

This data was developed using ab62373, the same antibody clone in a different buffer formulation.

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-beta Dystroglycan antibody [EP2200Y] (<a href='/en-us/products/primary-antibodies/beta-dystroglycan-antibody-ep2200y-ab62373'>ab62373</a>) at 1/1000 dilution

All lanes:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 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: 97 kDa

Observed band size: 43 kDa

false