Revision 1

#9932Store at -20C

1 Kit

(8 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
CDK2 (78B2) Rabbit mAb 2546 20 µl 33 kDa Rabbit 
p27 Kip1 (D69C12) XP® Rabbit mAb 3686 20 µl 27 kDa Rabbit IgG
Cyclin D1 (92G2) Rabbit mAb 2978 20 µl 36 kDa Rabbit IgG
CDK6 (DCS83) Mouse mAb 3136 20 µl 36 kDa Mouse IgG1
Cyclin D3 (DCS22) Mouse mAb 2936 20 µl 31 kDa Mouse IgG1
p21 Waf1/Cip1 (12D1) Rabbit mAb 2947 20 µl 21 kDa Rabbit IgG
CDK4 (D9G3E) Rabbit mAb 12790 20 µl 30 kDa Rabbit IgG
p18 INK4C (DCS118) Mouse mAb 2896 20 µl 18 kDa Mouse IgG2a
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

Cell Cycle Regulation Antibody Sampler kit offers an economical way of detecting eight integral cell cycle regulation proteins. The kit contains enough primary and secondary antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Eukaryotic cell cycle progression is dependent, in part, on the tightly regulated activity of cyclin dependent kinases (CDKs). Cyclin D/CDK4/6 activity occurs in mid-late G1 phase, upstream of CDK2/cyclin E activity. Both of these activities are required for hyperphosphorylation of the retinoblastoma gene product (pRb). pRb phosphorylation allows the release of S phase-promoting transcription factors and is indicative of the cell's commitment to proliferate. This point in the cell cycle is known as the restriction point. Cyclin protein levels oscillate throughout the cell cycle, and their availability is a means of controlling CDK activity and cell proliferation. Cyclin D is degraded through the ubiquitin proteasome pathway in the absence of mitogenic signaling. Ubiquitination of cyclin D1 is enhanced by phosphorylation at Thr286 by glycogen synthase kinase 3b (GSK-3b) (1). p27/Kip1, p57 Kip2 and p21 Waf1/Cip1 are members of the Cip/Kip family of cyclin-dependent kinase inhibitors. They form heterotrimeric complexes with cyclins and CDKs, inhibiting kinase activity and blocking progression through G1/S phase (2). However, p21 may enhance assembly and activity of cyclin D/CDK4/6 complexes (3). Levels of p21 and p27 protein are controlled through ubiquitination and proteasomal degradation (4). Levels of p27 are upregulated in quiescent cells and in cells treated with negative cell cycle regulators. p27 nuclear localization is controlled by Akt-dependent phosphorylation at Thr157 (5). The inhibitors of CDK4 (INK4) family include p15 INK4B, p16 INK4A, p18 INK4C, and p19 INK4D. All INK4 proteins selectively inhibit CDK4/6 activity, either in a binary complex, or in a ternary complex including cyclin D, resulting in inhibition of cell division (6,7).

  1. Diehl, J.A. et al. (1997) Genes Dev 11, 957-72.
  2. Pestell, R.G. et al. (1999) Endocr Rev 20, 501-34.
  3. Cheng, M. et al. (1999) EMBO J 18, 1571-83.
  4. Sheaff, R.J. et al. (2000) Mol Cell 5, 403-10.
  5. Shin, I. et al. (2002) Nat Med 8, 1145-52.
  6. Guan, K.L. et al. (1994) Genes Dev 8, 2939-52.
  7. Hirai, H. et al. (1995) Mol Cell Biol 15, 2672-81.

Background References

    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.

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