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9870
Cell Cycle Regulation Antibody Sampler Kit II
Primary Antibodies
Antibody Sampler Kit

Cell Cycle Regulation Antibody Sampler Kit II #9870

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Cell Cycle Regulation Antibody Sampler Kit II: Image 1

Flow cytometric analysis of Jurkat cells, using Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb versus propidium iodide (DNA content).

Cell Cycle Regulation Antibody Sampler Kit II: Image 2

Western analysis of extracts from Hela cells that were untreated, treated with doxorubicin (0.5 µM, 24 hours), or with nocodazole (50 ng/ml, 24 hours), using Cyclin A2 (BF683) Mouse mAb.

Cell Cycle Regulation Antibody Sampler Kit II: Image 3

Flow cytometric analysis of Jurkat cells using Cyclin B1 (D5C10) XP® Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 (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 II: Image 4

Western blot analysis of extracts from MCF-7, SK-N-MC and HeLa cells, untreated or treated with the proteasome inhibitor MG-132, using Cyclin E2 Antibody.

Cell Cycle Regulation Antibody Sampler Kit II: Image 5

Flow cytometric analysis of Jurkat cells using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb versus propidium iodide (DNA content). The boxed population indicates Phospho-Histone H3 (Ser10) positive cells.

Cell Cycle Regulation Antibody Sampler Kit II: Image 6

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear and cytoplasmic localization using Myt1 Antibody.

Cell Cycle Regulation Antibody Sampler Kit II: 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 II: 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 II: Image 9

Immunoprecipitation of phospho-wee1 from A431 cells, untreated or EGF-treated, using Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb followed by Western blot using the same antibody.

Cell Cycle Regulation Antibody Sampler Kit II: Image 10

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 II: Image 11

Confocal immunofluorescent analysis of asynchronous HeLa cells labeled with Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb (green) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (red).

Cell Cycle Regulation Antibody Sampler Kit II: Image 12

Confocal immunofluorescent analysis of HT-29 cells using Cyclin B1 (D5C10) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Cell Cycle Regulation Antibody Sampler Kit II: Image 13

Confocal immunofluorescent analysis of HeLa cells using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Cell Cycle Regulation Antibody Sampler Kit II: Image 14

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Myt1 Antibody.

Cell Cycle Regulation Antibody Sampler Kit II: Image 15

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

Cell Cycle Regulation Antibody Sampler Kit II: Image 16

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 II: Image 17

Western blot analysis of extracts from A431 and H441 cells, untreated or EGF-treated, using Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb (upper) or Wee1 Antibody #4936 (lower).

Cell Cycle Regulation Antibody Sampler Kit II: Image 18

Western blot analysis of extracts from C6 cells, untreated (-) or treated with Nocodazole (0.1 μg/ml, 18 hr; +), using Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb (upper) or cdc2 Antibody #77055 (lower).

Cell Cycle Regulation Antibody Sampler Kit II: Image 19

Western blot analysis of extracts from various cell lines using Cyclin B1 (D5C10) XP® Rabbit mAb.

Cell Cycle Regulation Antibody Sampler Kit II: Image 20

Western blot analysis of extracts from HeLa cells, either untreated or treated with nocodazole (100 ng/ml for 18 hours), using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (upper) or Histone H3 Antibody #9715 (lower). Phospho-specificity of the antibody is shown by further treatment of the lysate with λ phosphatase.

Cell Cycle Regulation Antibody Sampler Kit II: Image 21

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Myt1 Antibody.

Cell Cycle Regulation Antibody Sampler Kit II: Image 22

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 II: 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).

Cell Cycle Regulation Antibody Sampler Kit II: Image 24

Western blot analysis of extracts from HeLa cells, untreated or hydroxyurea treated for 20 hours, using Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb.

Cell Cycle Regulation Antibody Sampler Kit II: Image 25

Western blot analysis of extracts from HT-29 cells, synchronized in S-phase by double thymidine block (2 nM, 16 hr) followed by release into fresh media for the indicated time, using Cyclin B1 (D5C10) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Cell Cycle Regulation Antibody Sampler Kit II: Image 26

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Myt1 Antibody in the presence of control peptide (left) or antigen specific peptide (right).

Cell Cycle Regulation Antibody Sampler Kit II: 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 II: Image 28

Western blot analysis of extracts from HT29 cells, untreated (lane 1), nocodazole-treated (50 ng/ml, lane 2), or UV-treated (lane 3), using Myt1 Antibody.

Cell Cycle Regulation Antibody Sampler Kit II: Image 29

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.

To Purchase # 9870T
Product # Size Price
9870T
1 Kit  (8 x 20 µl) $ 554

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb 4539 20 µl
  • WB
  • IP
  • IF
  • F
H M R Mk 34 Rabbit 
Cyclin A2 (BF683) Mouse mAb 4656 20 µl
  • WB
H 55 Mouse IgE
Cyclin B1 (D5C10) XP® Rabbit mAb 12231 20 µl
  • WB
  • IP
  • IF
  • F
H R 55 Rabbit IgG
Cyclin E2 Antibody 4132 20 µl
  • WB
H 48 Rabbit 
Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb 3377 20 µl
  • WB
  • IF
  • F
H M R Mk Z 17 Rabbit IgG
Myt1 Antibody 4282 20 µl
  • WB
  • IHC
H M R 60 to 70 Rabbit 
p21 Waf1/Cip1 (12D1) Rabbit mAb 2947 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 21 Rabbit IgG
Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb 4910 20 µl
  • WB
  • IP
H 95 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

Product Description

The Cell Cycle Regulation Sampler Kit II provides an economical means of evaluating cell cycle proteins. The kit contains enough primary and secondary antibodies to perform two western blot experiments.

Specificity / Sensitivity

Each antibody in the Cell Cycle Regulation Sampler Kit II detects endogenous levels of its target protein and does not typically cross react with other family members. Cyclin B1 (D5C10) XP® Rabbit mAb recognizes endogenous levels of total cyclin B1 protein. This antibody also detects a 100 kDa protein of unknown origin in some cell lines. Activation state antibodies recognize target proteins only when phosphorylated at the indicated residue. Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb detects endogenous levels of cdc2 protein only when phosphorylated at tyrosine 15. Based on sequence similarity, the antibody may cross-react with CDK2 and CDK3.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the carboxy-terminus of human cyclin E2 or the middle of mouse and human Myt1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues near the carboxy-terminus of human p21, residues near the amino terminus of human cyclin B1 protein, or recombinant human cyclin A2 protein. Activation state monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser10 of human histone H3, residues surrounding Ser642 of human Wee1, or residues surrounding Tyr15 of human cdc2.

Background

The critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Tyr15 and Thr14 (1). Phosphorylation of cdc2 by the protein kinases Wee1 and Myt1 at Thr14 and Tyr15 results in inhibition of cdc2 (2,3). Progression through the G1/S checkpoint and initiation of DNA replication requires cyclin E; traversing the G2/M checkpoint to initiate mitosis requires cyclin B, and cyclin A may be required for both S-phase and M-phase (4). The versatile p21 cyclin-dependent kinase inhibitor, which interacts with several cyclin-CDK complexes to regulate cyclin-CDK during the cell cycle, is regulated by phosphorylation and ubiquitin-mediated degradation (5). Phosphorylation of histone H3 at Ser10 and neighboring residues correlates with chromosomal condensation, which is essential for segregation of chromosomes during mitosis (6).

  1. Norbury, C. et al. (1991) EMBO J 10, 3321-9.
  2. McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
  3. Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.
  4. Pagano, M. et al. (1992) EMBO. J. 11, 961-71.
  5. Abbas, T. and Dutta, A. (2009) Nat Rev Cancer 9, 400-14.
  6. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.

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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