Anti-ATP10B antibody [MJF-D29757-57]

• WB

Lab

Western blot - Anti-ATP10B antibody [MJF-D29757-57] (AB325638)

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

In Western blot, ab325638 was shown to bind specifically to ATP10B. Target of interest was observed at 180 kDa in wild-type LS513 cell lysates (lane 1) with no signal observed at this size in ATP10B knockout cell line (lane 2) (ab324258).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) 1 : 10000 (124Kda).

All lanes:

Western blot - Anti-ATP10B antibody [MJF-D29757-57] (ab325638) at 1/1000 dilution

Lane 1:

LS513 (human Large intestine Cecum epithelial cell) whole cell lysate at 40 µg

Lane 2:

Western blot - Human ATP10B Knockout LS513 cell line (<a href='/en-us/products/cell-lines/human-atp10b-knockout-ls513-cell-line-ab324258'>ab324258</a>) at 40 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: 180 kDa,124 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-ATP10B antibody [MJF-D29757-57] (AB325638)

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

Negative control : HepG2, 293T

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) 1 : 10000 (124Kda).

All lanes:

Western blot - Anti-ATP10B antibody [MJF-D29757-57] (ab325638) at 1/1000 dilution

Lane 1:

SW1116 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 40 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: 180 kDa,124 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-ATP10B antibody [MJF-D29757-57] (AB325638)

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

This antibody does not cross-react with recombinant human ATP10A or ATP10D by western blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (180KDa).

All lanes:

Western blot - Anti-ATP10B antibody [MJF-D29757-57] (ab325638) at 1/1000 dilution

Lane 1:

293T(human embryonic kidney epithelial cell) transfected with a human ATP10B expression vector containing a His-tag, whole cell lysate at 10 µg

Lane 2:

293T transfected with a human ATP10A expression vector containing a His-tag, whole cell lysate at 40 µg

Lane 3:

293T transfected with a human ATP10D expression vector containing a His-tag, whole cell lysate at 2 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: 180 kDa

false

Exposure time: 26s