重组Anti-CK1 epsilon抗体[1/Casein Kinase Ie]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (AB302638)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CK1 epsilon with ab302638 at 1/100 9.9ug/ml dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mainly cytoplasmic staining in HeLa cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution (Red).

The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab302638 at a 1/100 dilution followed by ab150080 at a 1/500 dilution.

-ve control 2 : ab179513 at a 1/200 dilution followed by ab150117 at a 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (AB302638)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma small irregularly shaped cells) cells labelling CK1 epsilon with ab302638 at 1/100 9.9ug/ml dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mainly cytoplasmic staining in PC-12 cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution (Red).

. The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab302638 at a 1/100 dilution followed by ab150080 at a 1/500 dilution.

-ve control 2 : ab179513 at a 1/200 dilution followed by ab150117 at a 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (AB302638)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CK1 epsilon with ab302638 at 1/100 9.9ug/ml dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mainly cytoplasmic staining in NIH/3T3 cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution (Red).

. The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab302638 at a 1/100 dilution followed by ab150080 at a 1/500 dilution.

-ve control 2 : ab179513 at a 1/200 dilution followed by ab150117 at a 1/1000 dilution.

• WB

Lab

Western blot - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (AB302638)

Western blot : Anti-CK1 epsilon antibody [1/Casein Kinase Iepsilon] ab302638 staining at 1/1000 dilution, shown in green; Rabbit anti-ACTN4 ab108198 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 47 kDa in Wild-type A549 cell lysates with no signal observed at this size in CSNK1E knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW and Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (ab302638) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

CSNK1E knockout A549 at 20 µg

Lane 3:

Jurkat at 20 µg

Lane 4:

PC-3 at 20 µg

Secondary

All lanes:

Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa,110 kDa

false

• WB

Supplier Data

Western blot - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (AB302638)

WB also tested the KO lysated side by side with published Rabmab (ab270997), please find the in internal WB. Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 3 minutes

All lanes:

Western blot - Anti-CK1 epsilon antibody [1/Casein Kinase Iε] (ab302638) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell), whole cell lysate

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate

Lane 4:

C2C12 (mouse myoblasts), whole cell lysate

Lane 5:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate

Lane 7:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

false

Exposure time: 3min