#P06493
983
Product Information
Product Usage Information
As controls, CST recommends using 10 µl of GST-cdc2 negative control protein and 20 µl of the positive control SK-N-MC cell extract.
Note: Boil both negative control and positive control for 2-4 minutes prior to loading on an SDS-PAGE gel.
Storage
Product Description
Nonphosphorylated cdc2 Control Protein: A glutathione S-transferase (GST)-cdc2 fusion protein serves as a negative control for monitoring antibody phosphoselectivity for Western blotting experiments. Molecular weight of GST-cdc2 fusion protein is 32 kDa.
| MW (kDa) | 32, 34 |
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).
Description:
Phosphorylated cdc2 Control Protein: SK-N-MC cell extracts treated with hydroxyurea for 22 hours to induce cdc2 (Tyr15) phosphorylation serve as a positive control.
Nonphosphorylated cdc2 Control Protein: A glutathione S-transferase (GST)-cdc2 fusion protein serves as a negative control. The molecular weight of GST-cdc2 fusion protein is 32 kDa.
Western Blots: CST recommends using 10 µl of GST-cdc2 negative control protein and 20 µl of the positive control SK-N-MC cell extract.
Note: Boil both negative control and positive control for 2-4 minutes prior to loading on an SDS-PAGE gel.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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