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Western blot analysis of extracts from Saos cells, either untreated or treated with hydroxyurea or nocodazole, using Phospho-cdc2 (Tyr15) Antibody #9111.

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Western blot analysis of extracts from HeLa cells synchronized at various stages of the cell cycle, using cdc2 (POH1) Mouse mAb.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

Learn more about how we get our images

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Learn more about how we get our images

Confocal immunofluorescent analysis of HT-29 cells, transfected with SignalSilence® Control siRNA (Unconjugated) #6568 (left), SignalSilence® cdc2 siRNA I #3500 (center) or SignalSilence® cdc2 siRNA II #3600 (right), using cdc2 (POH1) Mouse mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Image
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-cdc2 (Tyr15) Antibody 9111 100 µl
H M R Mk Dm X Sc 34 Rabbit 
cdc2 (POH1) Mouse mAb 9116 100 µl
H Mk 34 Mouse IgG2a
cdc2 (Tyr15) Control Proteins 9113 100 µl
32, 34  
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
All Goat 
Anti-biotin, HRP-linked Antibody 7075 100 µl
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
All Horse 
Biotinylated Protein Ladder Detection Pack 7727 100 µl
 
20X LumiGLO® Reagent and 20X Peroxide 7003 5 ml each
 

Product Description

PhosphoPlus® Cdc2 (Tyr15) Western Detection Kit offers an efficient way of detecting cdc2 (Tyr15) phosphorylation by western blotting. The kit contains enough primary and secondary antibodies to perform 10 western blots, as well as a set of pre-stained and biotinylated markers, control proteins and LumiGLO® reagent.


Specificity / Sensitivity

When used in conjunction with our Phototope®-HRP Western Detection Kit, Phospho-cdc2 (Tyr15) Antibody detects less than 10 ng of tyrosine phosphorylated cdc2 yet will not react with up to 1 µg of non-tyrosine phosphorylated cdc2. Similarly, Phospho-cdc2 (Tyr15) Antibody detects phosphorylated tyrosine 15 of cdk2 but demonstrates no cross reactivity to cdk4, cdk6 and cdk7 or other phospho-tyrosine proteins.


Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the phsphorylation site. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a recombinant human cdc2 fusion protein.

The entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Thr14 and Tyr15 (2). Phosphorylation at Thr14 and Tyr15, resulting in inhibition of cdc2, can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).


1.  Hunter, T. (1995) Cell 80, 225-236.

2.  Norbury, C. et al. (1991) EMBO J 10, 3321-9.

3.  Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.

4.  McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.

5.  Atherton-Fessler, S. et al. (1994) Mol Biol Cell 5, 989-1001.


Entrez-Gene Id 983
Swiss-Prot Acc. P06493


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.