Anti-SHARPIN antibody [EPR29120-74]

• WB

Supplier Data

Western blot - Anti-SHARPIN antibody [EPR29120-74] (AB321891)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32165044).

The identity of the lower MW band at approximately 30 kDa (in lanes 1, 3 and 5) is unknown.

Exposure time : Lanes 1-5 : 147 seconds; Lane 6 : 8 seconds

All lanes:

Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891) at 1/1000 dilution

Lane 1:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

CT26.WT (mouse colon 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

Observed band size: 40 kDa

false

• WB

Supplier Data

Western blot - Anti-SHARPIN antibody [EPR29120-74] (AB321891)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32165044).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

THP-1 transfected with siRNA specifically targeting NTHP-1 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 1/100000 dilution

Observed band size: 40 kDa,36 kDa

true

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-SHARPIN antibody [EPR29120-74] (AB321891)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32165044).

To minimize protein degradation, cells (lanes 7-9) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Observed band size: 40 kDa

false

Exposure time: 180s