Product Pathways - Cell Cycle / Checkpoint

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Cell Cycle Regulation Antibody Sampler Kit #9932

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
CDK2 (78B2) Rabbit mAb #2546	40 µl	W IP F	H M R Mk	33	Rabbit IgG
Cyclin D1 (DCS6) Mouse mAb #2926	40 µl	W IP	H M R	36	Mouse IgG2a
p27 Kip1 Antibody #2552	40 µl	W IP	H M R	27	Rabbit
CDK6 (DCS83) Mouse mAb #3136	40 µl	W	H M R	36	Mouse IgG1
Cyclin D3 (DCS22) Mouse mAb #2936	40 µl	W IHC-P	H M R	31	Mouse IgG1
p21 Waf1/Cip1 (12D1) Rabbit mAb #2947	40 µl	W IP IHC-P IF-IC F	H Mk (Dg)	21	Rabbit IgG
CDK4 (DCS156) Mouse mAb #2906	40 µl	W	H M	30	Mouse IgG1
p18 INK4C (DCS118) Mouse mAb #2896	40 µl	W IP	H	18	Mouse IgG2a
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat
Anti-mouse IgG, HRP-linked Antibody #7076	100 µl				Horse

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Antibodies detect endogenous levels of their respective proteins.

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 four Western blot experiments with each primary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides and are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant human proteins or synthetic peptides.

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, 957-972.
2. Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534.
3. Cheng, J. et al. (1999) EMBO J. 18, 1571-1573.
4. Sheaff, R.J. et al. (2000) Cell 5, 403-410.
5. Shin, I. et al. (2002) Nat. Med. 8, 1145-1152.
6. Guan, K.L. et al. (1994) Genes Dev. 8, 2939-2952.
7. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.

Application References

• Chang, S.F. et al. (2008) Proc Natl Acad Sci U S A 105, 3927-32. Applications: Western Blotting
• Yamanaka, Y. et al. (2009) Blood 114, 3265-75. Applications: Western Blotting

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

2546:

Flow, Immunoprecipitation, Western Blotting

2552:

Immunoprecipitation, Western Blotting

2896:

Immunoprecipitation, Western Blotting

2906:

Western Blotting

2926:

Immunoprecipitation, Western Blotting

2936:

IHC / Paraffin, Western Blotting

2947:

Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

3136:

Western Blotting

Companion Products

• 9917 Cell Cycle/Checkpoint Antibody Sampler Kit
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 2922 Cyclin D1 Antibody
• 2924 Cyclin D2 Antibody
• 2896 p18 INK4C (DCS118) Mouse mAb

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For Research Use Only. Not For Use In Diagnostic Procedures.