Immunofluorescence analysis of CSNK1E / Casein Kinase I epsilon was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CSNK1E / Casein Kinase I epsilon Rabbit Polyclonal Antibody (Product # 48-7600) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the N-terminal region of the human casein kinase 1 epsilon protein (Accession# NP_689407), which is identical to mouse and rat sequence.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Casein kinases are a group of ubiquitous Ser/Thr kinases that phosphorylate key regulatory proteins involved in the control of cell differentiation, proliferation, chromosome segregation, circadian rhythms, and metabolic pathways. CK1 family members share a highly conserved kinase domain but differ in their variable N-and C-terminal domains. Mammals have seven family members: alpha, beta, gamma1, gamma2, gamma3, delta and epsilon. They all share at least 50% amino acid identity within the protein kinase catalytic domain. CK1 isoforms exclusively use ATP as phosphate donor and are generally co-factor independent. Casein kinase I epsilon is a Wnt-regulated kinase, and regulated phosphorylation of Dvl allows fine tuning of the Wnt-b-catenin signaling pathway. CKIe positively regulates Akt possibly by inhibiting PP2A. CKIe play an important role in neurodegenerative diseases.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.