Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

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.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using CDK2 (78B2) Rabbit mAb #2546.

Western Blotting

Western Blotting

Western blot analysis of whole cell extracts from C6, NIH/3T3, HeLa and MCF-7 cells using p27 Kip1 Antibody #2552.

Western Blotting

Western Blotting

Western blot analysis of SK-N-MC and IMCD3 cell lysates using CDK4 (DCS156) mAb #2906.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells using Cyclin D1 (DCS) mAb #2926.

Western Blotting

Western Blotting

Western blot analysis of extracts from SK-N-MC, C6 and IMCD3 cells using Cyclin D3 (DCS22) mAb #2936.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, IM-CD-3 and C6 cell lysates using CDK6 (DCS83) mAb #3136.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of cyclin D1 in paraffin-embedded human breast carcinoma showing nuclear localization, using Cyclin D1 (DCS6) mAb #2926.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of cyclin D3 in paraffin-embedded human breast carcinoma showing nuclear localization, using Cyclin D3 (DCS22) mAb #2936.


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

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

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