Immunofluorescence analysis of VRK1 was performed using 70% confluent log phase HeLa 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 VRK1 (1F6) Mouse Monoclonal Antibody (Product # MA1-828) at 1:100 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant fusion protein GST-VRK1.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (IHC)||1:100 - 1:500|
|Western Blot (WB)||1:500|
* 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.
MA1-828 detects VRK1 from human and mouse samples.
MA1-828 has been successfully used in Western blot, immunofluorescence, and immunohistochemistry applications. For Western blotting, it is recommended to block with 3% BSA (not milk).
The MA1-828 immunogen is recombinant fusion protein GST-VRK1.
Human vaccinia-related kinase 1 (VRK1) is a new kinase related to a poxvirus kinase, and more distantly to the casein kinase 1 family. VRK1 shows strong autophosphorylating activity in several Ser and Thr residues. VRK has also been found to be an upstream regulator of p53 that belongs to a new signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.