Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) untreated HeLa treated with 400 ng/ml Nocodazole for 20 hours HeLa treated with 400 ng/ml Nocodazole for 20 hours then treated with λ protein phosphatase treatment cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/5000 (0.096 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 positive staining in HeLa cells (shown in green) treated with 400 ng/ml nocodazole for 20 hours, the signal decreased after λ Protein phosphatase treatment at 30℃ for 2 hours . 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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) untreated A431 treated with 400 ng/ml Nocodazole for 20 hours A431treated with 400 ng/ml Nocodazole for 20 hours then treated with λ protein phosphatase treatment cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/5000 (0.096 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 positive staining in A431 cells (shown in green) treated with 400 ng/ml nocodazole for 20 hours, the signal decreased after λ Protein phosphatase treatment at 30℃ for 2 hours. 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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells transfected with an empty expression vector containing a myc-His-tag® 293T cells transfected with a human Aurora A (T288A) expression vector containing a myc-His-tag® 293T cells transfected with a human Aurora A (WT) expression vector containing a myc-His-tag® cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/5000 (0.096 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 positive staining in 293T cells (shown in green) transfected with a human Aurora A (WT) expression vector containing a myc-His-tag®.In contrast, no staining is observed in cells transfected with either the human Aurora A (T288A) mutant or an empty expression vector containing the same myc-His-tag®. 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).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 400 ng/ml nocodazole for 20 hours (Green) Untreated control (Magenta) cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/500 dilution (0.1ug) / Magenta and Green compared with a Rabbit monoclonal IgG (ab172730) / Black and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were treated with alkaline phosphatase over night (Right) / Untreated cells (Left)

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with DRAQ5 to differentiate cell cycle phase.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized L-929 (mouse connective tissue fibroblast) treated with 100 ng/mL Nocodazole for 14hours (Lower left and right) Untreated L-929 (Upper left and right) cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/5000 dilution (0.01ug) / Upper right and Lower right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with DRAQ5 to differentiate cell cycle phase.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized L-929 (mouse connective tissue fibroblast) untreated L-929 treated with 100 ng/ml Nocodazole for 14 hours L-929 treated with 100 ng/ml Nocodazole for 14 hours then treated with Alkaline phosphatase treatment cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/5000 (0.096 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 positive staining in L-929 cells (shown in green) treated with 100 ng/ml nocodazole for 14 hours, the signal decreased after Alkaline phosphatase treatment at 37℃ for 2 hours. 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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) untreated C6 treated with 100 ng/ml Nocodazole for 14 hours C6 treated with 100 ng/ml Nocodazole for 14 hours then treated with Alkaline phosphatase treatment cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/5000 (0.096 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 positive staining in C6 cells (shown in green) treated with 100 ng/ml nocodazole for 14 hours, the signal decreased after Alkaline phosphatase treatment at 37℃ for 2 hours. 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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) treated with 100 ng/mL Nocodazole for 14hours (Lower left and right) Untreated C6 (Upper left and right) cells labelling Aurora kinase A (phospho T288) with ab324635 at 1/50 dilution (1ug) / Upper right and Lower right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with DRAQ5 to differentiate cell cycle phase.

• Dot

Lab

Dot Blot - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (AB324635)

Dot blot analysis of Aurora kinase A (phospho T288) using ab324635 at 1000 (0.48 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : Aurora kinase A (phospho T288) peptide a

Lane 2 : Aurora kinase A (phospho T288) peptide b

Lane 3 : Aurora kinase A non-phospho peptide c

Lane 4 : Aurora kinase A (phospho T287) peptide d

Lane 5 : Aurora kinase A (phospho T232) peptide e

Exposure time : 180 seconds.

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

All lanes:

Dot Blot - Anti-Aurora kinase A (phospho T288) antibody [EPR28759-80] (ab324635) at 1/1000 dilution

Lane 1:

Aurora kinase A (phospho T288) peptide a

Lane 2:

Aurora kinase A (phospho T288) peptide b

Lane 3:

Aurora kinase A non-phospho peptide c

Lane 4:

Aurora kinase A (phospho T287) peptide d

Lane 5:

Aurora kinase A (phospho T232) peptide e

Secondary

All lanes:

Dot 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

false

Exposure time: 180s