Anti-Shootin-1 antibody [EPR29089-44]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Shootin-1 antibody [EPR29089-44] (AB322653)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized SK-OV-3(human ovarian cancer epithelial cell) cells labelling Shootin-1 with ab322653 at 1/100 (5.23 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in SK-OV-3 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : Daudi.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.

• WB

Supplier Data

Western blot - Anti-Shootin-1 antibody [EPR29089-44] (AB322653)

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

Low expression : Daudi.

The identity of the lower MW band at approximately 25 kDa is unknown.

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

All lanes:

Western blot - Anti-Shootin-1 antibody [EPR29089-44] (ab322653) at 1/1000 dilution

Lane 1:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 3:

Daudi (human Burkitts lymphoma lymphoblast) 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: 53 kDa,76 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Shootin-1 antibody [EPR29089-44] (AB322653)

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

Low expression : Mouse adult brain (PMID : 26849555, PMID : 17030985).

Negative control : Mouse spleen, mouse heart (PMID : 17030985; PMID : 27177867).

The identity of the higher MW band at approximately 250 kDa is unknown.

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

All lanes:

Western blot - Anti-Shootin-1 antibody [EPR29089-44] (ab322653) at 1/1000 dilution

Lane 1:

Mouse P0 brain tissue lysate at 20 µg

Lane 2:

Mouse P0 hippocampus tissue lysate at 20 µg

Lane 3:

Mouse adult brain tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Mouse heart tissue lysate at 20 µg

Lane 6:

Rat P5 brain tissue lysate at 20 µg

Lane 7:

Rat brain tissue lysate at 20 µg

Lane 8:

Rat spleen tissue lysate at 20 µg

Lane 9:

Rat heart tissue 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: 53 kDa,76 kDa

false

Exposure time: 26s