Anti-TMEM41B antibody [EPR28149-41] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-TMEM41B antibody [EPR28149-41] - BSA and Azide free (AB314764)

This data was developed using ab314763, the same antibody clone in a different buffer formulation. TMEM41B was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab314763 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314763 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate Lane 2 : ab314763 IP in 293T (human embryonic kidney epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314763 in 293T whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TMEM41B antibody [EPR28149-41] (<a href='/en-us/products/primary-antibodies/tmem41b-antibody-epr28149-41-ab314763'>ab314763</a>) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-TMEM41B antibody [EPR28149-41] - BSA and Azide free (AB314764)

This data was developed using ab314763, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : Lane 1 : 180 seconds Lane 2 : 59 seconds Lane 3 : 26 seconds

All lanes:

Western blot - Anti-TMEM41B antibody [EPR28149-41] (<a href='/en-us/products/primary-antibodies/tmem41b-antibody-epr28149-41-ab314763'>ab314763</a>) at 1/1000 dilution

Lane 1:

NCI-H1299 (human lu carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HCT-15 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) 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

Observed band size: 32 kDa

false

• WB

Supplier Data

Western blot - Anti-TMEM41B antibody [EPR28149-41] - BSA and Azide free (AB314764)

This data was developed using ab314763, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-TMEM41B antibody [EPR28149-41] (<a href='/en-us/products/primary-antibodies/tmem41b-antibody-epr28149-41-ab314763'>ab314763</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNAs specifically targeting TMEM41B 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

Observed band size: 32 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-TMEM41B antibody [EPR28149-41] - BSA and Azide free (AB314764)

This data was developed using ab314763, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EPR28149-41] to TMEM41B ab314763 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 25 kDa in Wild-type A549 cell lysates with no signal observed at this size in TMEM41B knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TMEM41B antibody [EPR28149-41] (<a href='/en-us/products/primary-antibodies/tmem41b-antibody-epr28149-41-ab314763'>ab314763</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

TMEM41B knockout A549 cell line at 20 µg

Lane 3:

JEG-3 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

false