Anti-MSH3 antibody [EPR29050-69]

• WB

Supplier Data

Western blot - Anti-MSH3 antibody [EPR29050-69] (AB324225)

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

To minimize protein degradation, cells (lanes 2-7) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

Lanes 1 is incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 2-7 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000.

The identity of the band at approximately 36 kDa in lanes 2-7 is unknown.

The identity of the band at approximately 15 kDa in lanes 1 is unknown.

The identity of the band at approximately 250 kDa in lanes 2-3 is unknown.

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

Exposure time : Lanes 1, Lanes 4-7 : 180 seconds; Lanes 2-3 : 48 seconds

All lanes:

Western blot - Anti-MSH3 antibody [EPR29050-69] (ab324225) at 1/1000 dilution

Lane 1:

Human pancreas cancer tissue lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Mouse testis tissue lysate at 20 µg

Lane 7:

Mouse spleen tissue lysate at 20 µg

Secondary

Lane 1:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 2 - 7:

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: 120 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-MSH3 antibody [EPR29050-69] (AB324225)

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

Performed under reducing conditions.

In Western blot, ab324225 was shown to bind specifically to MSH3. Target of interest was observed at 120 kDa in wild-type A549 cell lysates (lane 1) with no signal observed at this size in MSH3 knockout cell line (lane 2) (lane 2, knockout cell line ab288875).

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

All lanes:

Western blot - Anti-MSH3 antibody [EPR29050-69] (ab324225) at 1/1000 dilution

Lane 1:

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

Lane 2:

MSH3 knockout A549 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: 120 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MSH3 antibody [EPR29050-69] (AB324225)

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

The identity of the band at approximately 36 kDa is unknown.

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

All lanes:

Western blot - Anti-MSH3 antibody [EPR29050-69] (ab324225) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 3:

NIH/3T3 transfected with siRNA specifically targeting MSH3 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: 120 kDa,36 kDa

false

Exposure time: 26s