Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

PhosphoPlus® cdc2 (Tyr15) Antibody Kit #9110

Kit Includes Quantity Applications Reactivity MW (kDa) Source
Phospho-cdc2 (Tyr15) Antibody # 9111 0.1 milliliters W IP E-P H M R Mk Sc Dm X 34 Rabbit
cdc2 Antibody # 9112 0.1 milliliters W H M R 34 Rabbit
cdc2 (Tyr15) Control Proteins # 9113 40 microliters 32, 34
Anti-rabbit IgG, HRP-linked Antibody # 7074 50 microliters Goat
Anti-biotin, HRP-linked Antibody # 7075 0.1 milliliters Goat
Biotinylated Protein Ladder Detection Pack # 7727 0.1 milliliters
20X LumiGLO® Reagent and 20X Peroxide # 7003 5 milliliters each
cdc2 (Tyr15) Control Proteins # 9113 80 microliters 32, 34

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  E-P=ELISA (Peptide)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Sc=S. cerevisiae  Dm=D. melanogaster  X=Xenopus

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.

Western Blotting

Western Blotting

Western analysis of extracts from hydroxyurea (G1/S) or nocodizate (G2/M) treated Saos cells with Phospho-cdc2 (Tyr15) (A) and cdc2 (Tyr15) Antibodies (B).

Table

Table

Role of cdc2 Y15 phosporylation in the progression of eukaryotic cells into M-phase of the cell cycle.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH coupled) corresponding to residues surrounding the phsphorylation site. Antibodies are purified by protein A and peptide affinity chromatography.

Background

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 Tyr15 and Thr14 (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. Atherton-Fessler, S. et al. (1994) Mol. Biol. Cell. 5, 989-1001.
  2. Norbury, C. et al. (1991) EMBO. J. 10, 3321-3329.
  3. McGowan, C.H. and Russell, P. (1993) EMBO J. 12, 75-85.
  4. Wells, N.J. et al. (1999) J. Cell. Sci. 112, 3361-3371.
  5. Hunter, T. (1995) Cell 80, 225-236.

Application References

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