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9932
Cell Cycle Regulation Antibody Sampler Kit

Cell Cycle Regulation Antibody Sampler Kit #9932

Western Blotting Image 1

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

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Western Blotting Image 2

Western blot analysis of extracts from various cell types using p27 Kip1 (D69C12) XP® Rabbit mAb.

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Western Blotting Image 3

Western blot analysis of extracts from MCF7, L929 and C6 cells, using Cyclin D1 (92G2) Rabbit mAb.

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Western Blotting Image 4

Western blot analysis of extracts from HeLa, IM-CD-3 and C6 cells, using CDK6 (DCS83) Mouse mAb.

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Western Blotting Image 5

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

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Western Blotting Image 6

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

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Western Blotting Image 7

Western blot analysis of extracts from various cell lines using CDK4 (D9G3E) Rabbit mAb.

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Western Blotting Image 8

Western blot analysis of extracts from HeLa, 293 and Ramos cells, using p18 INK4C (DCS118) Mouse mAb.

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Western Blotting Image 9

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Flow Cytometry Image 10

Flow cytometric analysis of Jurkat cells, using CDK2 (78B2) Rabbit mAb versus propidium iodide (DNA content).

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IP Image 11

Immunoprecipitation of p27 Kip1 from 293 cells using p27 Kip1 (D69C12) XP® Rabbit mAb. Western analysis was performed using the same antibody. Lane 1 is 5% input.

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Cyclin D1 (92G2) Rabbit mAb.

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Cyclin D3 (DCS22) Mouse mAb.

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Western Blotting Image 14

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p21 Waf1/Cip1 siRNA II (+), using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947 and α-Tubulin (11H10) Rabbit mAb #2125. The p21 Waf1/Cip1 (12D1) Rabbit mAb confirms silencing of p21 Waf1/Cip1 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p21 Waf1/Cip1 siRNA.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using CDK4 (D9G3E) Rabbit mAb.

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Flow Cytometry Image 16

Flow cytometric analysis of Jurkat cells using p27 Kip1 (D69C12) XP® Rabbit mAb versus Propidium Iodide (PI)/RNase Staining Solution #4087. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine using Cyclin D1 (92G2) Rabbit mAb.

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IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Cyclin D3 (DCS22) Mouse mAb.

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IP Image 19

Immunoprecipitation of p21 from human umbillical vein endothelial cells (HUVECs) using p21 Waf1/Cip1 (12D1) Rabbit mAb. Western blot detection was performed using the same antibody.

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IHC-P (paraffin) Image 20

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CDK4 (D9G3E) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

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IF-IC Image 21

Confocal immunofluorescent analysis of MCF-7 cells using p27 Kip1 (D69C12) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Cyclin D1 (92G2) Rabbit mAb in the presence of control peptide (left) or Cyclin D1 Blocking Peptide #1044 (right).

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IHC-P (paraffin) Image 23

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using p21 Waf1/Cip1 (12D1) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

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Flow Cytometry Image 24

Flow cytometric analysis of Jurkat cells using CDK4 (D9G3E) Rabbit mAb and Propidium Iodid (PI)/RNase Staining Solution #4087 to measure DNA content. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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IHC-P (paraffin) Image 25

Immunohistochemical analysis of paraffin-embedded H1975 xenograft, using Cyclin D1 (92G2) Rabbit mAb.

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IHC-P (paraffin) Image 26

Immunohistochemical analysis of paraffin-embedded HeLa cells, transfected with SignalSilence® Control siRNA (Unconjugated) #6568 (left) or SignalSilence® p21 Waf1/Cip1 siRNA II #6558 (right), using p21 Waf1/Cip1 (12D1) Rabbit mAb.

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IF-IC Image 27

Confocal immunofluorescent analysis of MCF7 cells using CDK4 (D9G3E) Rabbit mAb (green), p21 Waf1/Cip1 (12D1) Rabbit mAb (Alexa Fluor® 555 Conjugate) #8493 (red), and Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #3458 (blue pseudocolor).

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Flow Cytometry Image 28

Flow cytometric analysis of untreated HT29 cells, using Cyclin D1 (92G2) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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Flow Cytometry Image 29

Flow cytometric analysis of HeLa cells (red) and MCF7 cells (blue), using p21 Waf1/Cip1 (12D1) Rabbit mAb.

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IF-IC Image 30

Confocal immunofluorescent analysis of MCF7 cells using p21 Waf1/Cip1 (12D1) Rabbit mAb (red) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CDK2 (78B2) Rabbit mAb 2546 20 µl
  • WB
  • IP
  • F
H M R Mk 33 Rabbit 
p27 Kip1 (D69C12) XP® Rabbit mAb 3686 20 µl
  • WB
  • IP
  • IF
  • F
H R Mk 27 Rabbit IgG
Cyclin D1 (92G2) Rabbit mAb 2978 20 µl
  • WB
  • IHC
  • F
H M R 36 Rabbit IgG
CDK6 (DCS83) Mouse mAb 3136 20 µl
  • WB
H M R 36 Mouse IgG1
Cyclin D3 (DCS22) Mouse mAb 2936 20 µl
  • WB
  • IHC
H M R 31 Mouse IgG1
p21 Waf1/Cip1 (12D1) Rabbit mAb 2947 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 21 Rabbit IgG
CDK4 (D9G3E) Rabbit mAb 12790 20 µl
  • WB
  • IHC
  • IF
  • F
H Mk 30 Rabbit IgG
p18 INK4C (DCS118) Mouse mAb 2896 20 µl
  • WB
  • IP
H 18 Mouse IgG2a
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

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.

Antibodies detect endogenous levels of their respective proteins.

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.

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.
Entrez-Gene Id
1017 , 1019 , 1021 , 595 , 896 , 1031 , 1026 , 1027
Swiss-Prot Acc.
P24941 , P11802 , Q00534 , P24385 , P30281 , P42773 , P38936 , P46527
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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