Immunofluorescence analysis of Cdk5 was performed using 70% confluent log phase SH-SY5Y 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 Cdk5 (8A1) Mouse Monoclonal Antibody (LFMA0116) at 1:250 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 (A28175) at 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 (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Purified, His-tagged, recombinant, human CDK5 protein, from baculovirus expressed insect cells.|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunoprecipitation (IP)||2 µl|
|Western Blot (WB)||1:250|
* 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.
A suggested positive control for this product is SH-SY5Y cells.
Cyclin-dependent kinase-5 (CDK5) is a member of the cyclin-dependent kinase family of serine/threonine kinases. Its mRNA and protein are expressed in the kidneys, testes, and ovaries. And Its activity has been detected almost exclusively in brain extracts. Similar to other Cdks, monomeric Cdk5 displays no enzymatic activity; however, Cdk5 is not activated bycyclins. Instead, Cdk5 activity requires association with one of two brain-specific regulatory subunits, called p35 and p39. These two activators regulate the spatial and temporal expression of active Cdk5, restricting its activity primarily to post-mitotic neurons.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.