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9932
Cell Cycle Regulation Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Cell Cycle Regulation Antibody Sampler Kit #9932

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Cell Cycle Regulation Antibody Sampler Kit: Image 1

Flow cytometric analysis of Jurkat cells using CDK2 (78B2) Rabbit mAb (right) and Propidium Iodide (PI)/RNase Staining Solution #4087, compared to a secondary only sample (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Cell Cycle Regulation Antibody Sampler Kit: Image 2

Western blot analysis from control HeLa cells (lane 1) or p27 Kip1 knockout HeLa cells (lane 2) using p27 Kip1 (D69C12) XP Rabbit mAb #3686 (upper) or B-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the p27 Kip1 knockout HeLa cells confirms specificity of the antibody for p27 Kip1.

Cell Cycle Regulation Antibody Sampler Kit: Image 3

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 4

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

Cell Cycle Regulation Antibody Sampler Kit: Image 5

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

Cell Cycle Regulation Antibody Sampler Kit: Image 6

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

Cell Cycle Regulation Antibody Sampler Kit: Image 7

Western blot analysis from control HeLa cells (lane 1) or p21 Waf1/Cip1 knockout HeLa cells (lane 2) using p21 Waf1/Cip1 (12D1) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the p21 Waf1/Cip1 knockout HeLa cells confirms specificity of the antibody for p21 Waf1/Cip1.

Cell Cycle Regulation Antibody Sampler Kit: Image 8

Flow cytometric analysis of Daudi cells using p21 Waf1/Cip1 (12D1) Rabbit mAb (right) and Propidium Iodide (PI)/RNase Staining Solution #4087, compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Cell Cycle Regulation Antibody Sampler Kit: Image 9

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 10

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

Cell Cycle Regulation Antibody Sampler Kit: Image 11

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 12

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

Cell Cycle Regulation Antibody Sampler Kit: Image 13

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

Cell Cycle Regulation Antibody Sampler Kit: Image 14

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).

Cell Cycle Regulation Antibody Sampler Kit: Image 15

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

Cell Cycle Regulation Antibody Sampler Kit: Image 16

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

Cell Cycle Regulation Antibody Sampler Kit: Image 17

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

Cell Cycle Regulation Antibody Sampler Kit: Image 18

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).

Cell Cycle Regulation Antibody Sampler Kit: Image 19

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).

Cell Cycle Regulation Antibody Sampler Kit: Image 20

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 21

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).

Cell Cycle Regulation Antibody Sampler Kit: Image 22

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

Cell Cycle Regulation Antibody Sampler Kit: Image 23

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 24

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).

Cell Cycle Regulation Antibody Sampler Kit: Image 25

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

Cell Cycle Regulation Antibody Sampler Kit: Image 26

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

Cell Cycle Regulation Antibody Sampler Kit: Image 27

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 28

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).

Cell Cycle Regulation Antibody Sampler Kit: Image 29

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

Cell Cycle Regulation Antibody Sampler Kit: Image 30

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.

Cell Cycle Regulation Antibody Sampler Kit: Image 31

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

To Purchase # 9932T
Product # Size Price
9932T
1 Kit  (8 x 20 µl) $ 554

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
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 

Product 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.

Specificity / Sensitivity

Antibodies detect endogenous levels of their respective proteins.

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, 957-72.
  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.

Limited Uses

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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's 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's 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.

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