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Western blot analysis was performed on whole cell extracts (30 ug lysate) of HeLa (1), HeLa treated for 20 hr with 3mM of Hydroxy Urea (2), HeLa Serum Starved (3), HeLa Serum Starved overnight following by serum release (4), HeLa treated with UV for 40 min (5), Jurkat (6), Jurkat Serum Starved (7), Jurkat Serum Starved overnight following by serum release (8), Jurkat treated with UV for 40 min (9) and U2OS (10). The blots were probed with Anti-CDC2 [pT14/pY15] Rabbit Polyclonal Antibody (Product# 44686G, 1:500-1:2000 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Prod# G21234, 1:5000 dilution). A 34 kDa band corresponding to CDC2 [pT14/pY15] was observed across cell lines tested. Upon UV treatment and Serum starvation overnight followed by serum release, the expression of CDC2 [pT14/pY15] was decreased. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12% Bis-Tris gel (Prod# NP0342BOX), XCell SureLock™ Electrophoresis System (Prod# EI0002) and Novex® Sharp Pre-Stained Protein Standard (Prod# LC5800). Resolved proteins were then transferred onto a transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Prod# 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 (Prod# WP20005)
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Yeast, Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antibody was produced against a synthetic phosphopeptide derived from the region of the human cyclin-dependent kinase that contains threonine 14 and tyrosine 15.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:500-1:2000|
* 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.
|Western Blot (WB)||See 3 publications below|
CDK1 is a member of the Ser/Thr protein kinase family. This protein is a catalytic subunit of the highly conserved protein kinase complex known as M-phase promoting factor (MPF), which is essential for G1/S and G2/M phase transitions of eukaryotic cell cycle. Mitotic cyclins stably associate with this protein and function as regulatory subunits. The kinase activity of this protein is controlled by cyclin accumulation and destruction through the cell cycle. The phosphorylation and dephosphorylation of this protein also play important regulatory roles in cell cycle control.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A novel coumarin-quinone derivative SV37 inhibits CDC25 phosphatases, impairs proliferation, and induces cell death.
44-686G was used in western blot to characterize new inhibitors of CDC25.
|Bana E,Sibille E,Valente S,Cerella C,Chaimbault P,Kirsch G,Dicato M,Diederich M,Bagrel D||Molecular carcinogenesis (54:229)||2015|
Upregulation of p21 activates the intrinsic apoptotic pathway in β-cells.
44-686G was used in western blot to study the role of p21 in beta cell apoptosis
|Hernandez AM,Colvin ES,Chen YC,Geiss SL,Eller LE,Fueger PT||American journal of physiology. Endocrinology and metabolism (304:E1281)||2013|
|Yeast||Not Cited||Multisite phosphoregulation of Cdc25 activity refines the mitotic entrance and exit switches.||Lu LX,Domingo-Sananes MR,Huzarska M,Novak B,Gould KL||Proceedings of the National Academy of Sciences of the United States of America (109:9899)||2012|
CDC2; CDC28A; CDKN1; cell cycle controller CDC2; cell cycle p34 CDC2 kinase protein; cell division control protein 2 homolog; cell division cycle 2 homolog A; cell division cycle 2, G1 to S and G2 to M; Cell division cycle control protein 2; cell division cycle control protein 2a; cell division protein kinase 1; cyclin-dependent kinase 1; p34 protein kinase; P34CDC2
CDC2; CDC28A; Cdc2a; CDK1; CDKN1; p34<CDC2>; P34CDC2