WB, W-S, IP, IHC-P, FC-FP
H Mk
Endogenous
95
Rabbit IgG
#P30291
7465
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunoprecipitation | 1:50 |
| Immunohistochemistry (Paraffin) | 1:100 - 1:400 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 - 1:400 |
Storage
For a carrier free (BSA and azide free) version of this product see product #53692.
Specificity / Sensitivity
Species Reactivity:
Human, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala54 of human Wee1 protein.
Background
Entry of all eukaryotic cells into mitosis is regulated by activation of cdc2 kinase. The critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of Tyr15 and Thr14 (1,2). Phosphorylation at Tyr15 and Thr14 and inhibition of cdc2 is carried out by Wee1 and Myt1 protein kinases, while Tyr15 dephosphorylation and activation of cdc2 is carried out by the cdc25 phosphatase (1,3,4). Hyperphosphorylation and inactivation of Myt1 in mitosis suggests that one or more kinases activated at the G2/M transition negatively regulates Myt1 activity. Kinases shown to phosphorylate Myt1 include cdc2, p90RSK, Akt, and Plk1 (5-7).
Wee1 is inactivated upon mitotic entry by phosphorylation at Ser53 and Ser123 by Plk1 and cdc2, followed by β-TrCP-mediated ubiquitination and degradation (1,9,10).
- Watanabe, N. et al. (1995) EMBO J 14, 1878-91.
- Hunter, T. (1995) Cell 80, 225-36.
- Galaktionov, K. et al. (1995) Genes Dev 9, 1046-58.
- McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
- Booher, R.N. et al. (1997) J Biol Chem 272, 22300-6.
- Palmer, A. et al. (1998) EMBO J 17, 5037-47.
- Nakajima, H. et al. (2003) J Biol Chem 278, 25277-80.
- Parker, L.L. et al. (1995) Proc Natl Acad Sci U S A 92, 9638-42.
- Watanabe, N. et al. (2004) Proc Natl Acad Sci U S A 101, 4419-24.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)
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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