Western blot analysis was performed on whole cell extracts (30 ug lysate) of SH-SY5Y (Lane 1), PC-3 (Lane 2), HeLa (Lane 3), MCF7 (Lane 4), MDA-MB-231 (Lane 5), A549 (Lane 6) and Jurkat (Lane 7). The blot was probed with CDK5 Mouse monoclonal Antibody (Product# LF-MA0120, 1:250 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 ug/ml, 1:4000 dilution). A 33 kDa band corresponding to CDK5 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified, His-tagged, recombinant, human CDK5 protein expressed in E. coli.|
|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.