Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

PhosphoPlus® cdc2 (Tyr15) Antibody Kit #9110

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-cdc2 (Tyr15) Antibody #9111 100 µl W IP H M R Mk Dm X 34 Rabbit
cdc2 Antibody #9112 100 µl W H M R 34 Rabbit
cdc2 (Tyr15) Control Proteins #9113 100 µl 32, 34
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Anti-biotin, HRP-linked Antibody #7075 100 µl Goat
Biotinylated Protein Ladder Detection Pack #7727 100 µl
20X LumiGLO® Reagent and 20X Peroxide #7003 5 ml each

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dm=D. melanogaster  X=Xenopus
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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).

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.

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.

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 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. 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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