Anti-TAOK2 antibody [EPR30549-532]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TAOK2 antibody [EPR30549-532] (AB325603)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized wildtype HEK-293T(human embryonic kidney epithelial cell), ab255449;TAOK2 KO HEK-293T(TAOK2 knockout HEK-293T), ab266382; cells labelling TAOK2 with ab325603 at 1/500 (1.02 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in wildtype HEK-293T cells (shown in green), showing no staining in TAOK2 knockout HEK-293T cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 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 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TAOK2 antibody [EPR30549-532] (AB325603)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse splenocyte cells labelling TAOK2 with ab325603 at 1/2000 (0.255 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Low expression : Confocal image showing no staining in mouse splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 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 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TAOK2 antibody [EPR30549-532] (AB325603)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling TAOK2 with ab325603 at 1/2000 (0.255 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 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 dilution.

• WB

Lab

Western blot - Anti-TAOK2 antibody [EPR30549-532] (AB325603)

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

Low expression : human lymph node.

Performed under reducing conditions.

In Western blot, ab325603 was shown to bind specifically to TAOK2. Target of interest was observed at 150 and 200 kDa in wild-type 293T cell lysates (lane 1) with no signal observed at this size in TAOK2 knockout cell line (lane 2, knockout cell line ab266382).

The bands beneath the target band (150 kDa) are likely to be degraded target fragments.

The identity of the lower MW band at approximately 40 kDa (in lanes 1-2) is unknown.

Lanes 1-3 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 4-6 are incubated with Goat Anti-Rabbit IgG (HRP) at 1/2000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-TAOK2 antibody [EPR30549-532] (ab325603) at 1/1000 dilution

Lane 1:

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

Lane 2:

TAOK2 knockout 293T whole cell lysate at 20 µg

Lane 3:

HT-1376 (human urinary bladder carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Human hypothalamus tissue lysate at 20 µg

Lane 5:

Human cerebellum tissue lysate at 20 µg

Lane 6:

Human lymph node tissue lysate at 20 µg

Secondary

Lanes 1 - 3:

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

Lanes 4 - 6:

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

Observed band size: 150 kDa,200 kDa,36 kDa

false

Exposure time: 125s

• WB

Lab

Western blot - Anti-TAOK2 antibody [EPR30549-532] (AB325603)

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

Low expression : mouse lymph node.

The bands beneath the target band (150 kDa) are likely to be degraded target fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-TAOK2 antibody [EPR30549-532] (ab325603) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse lymph node tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 150 kDa,36 kDa

false

Exposure time: 26s