Revision 1

#5884Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

30

SOURCE:

Rabbit

UniProt ID:

#P11802

Entrez-Gene Id:

1019

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-CDK4 (Thr172) Antibody recognizes endogenous levels of CDK4 protein only when phosphorylated at Thr172.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Rat, Monkey, Dog, Pig

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr172 of human CDK4 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Cyclin-dependent kinase activity is regulated by T-loop phosphorylation (Thr172 in the case of CDK4), by the abundance of their cyclin partners, and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of CDK4/cyclin D and p27 Kip1/Cip1 requires extracellular mitogenic stimuli for the release and degradation of p27, which affects progression through the restriction point and pRb-dependent entry into S-phase (2). The active complex of CDK4/cyclin D targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). In HeLa cells, upon UV irradiation, upregulation of p16 INK4A association with CDK4/cyclin D3 leads to a G2 delay, implicating CDK4/cyclin D3 activity in progression through the G2-phase of the cell cycle (4).

  1. Hirai, H. et al. (1995) Mol Cell Biol 15, 2672-81.
  2. Sherr, C.J. (1996) Science 274, 1672-7.
  3. Lukas, J. et al. (1996) Mol Cell Biol 16, 6917-25.
  4. Gabrielli, B.G. et al. (1999) J Biol Chem 274, 13961-9.

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

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#5884

Phospho-CDK4 (Thr172) Antibody

Western Blotting Image 1: Phospho-CDK4 (Thr172) Antibody Expand Image
Western blot analysis of extracts from HT-29 cells, untreated or synchronized in G1 phase, using Phospho-CDK4 (Thr172) Antibody (upper) or CDK4 (DCS156) Mouse mAb #2906 (lower). After western transfer, the membrane was either left untreated (-CIP, left) or treated with Calf Intestinal Phosphatase (+CIP, right). G1 phase synchronization was performed by thymidine block (2 mM, 16 hours), release into nocodazole (100 ng/mL, 24 hours), selective detachment of mitotic cells, and release into G1 phase (4 hours).
Western Blotting Image 2: Phospho-CDK4 (Thr172) Antibody Expand Image
Western blot analysis of extracts from U-2 OS cells, untreated or synchronized in G1 phase, using Phospho-CDK4 (Thr172) Antibody. The antibody was preincubated with a phospho-CDK4 (Thr172) peptide or a nonphosphorylated CDK4 (Thr172) peptide.