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9870
Cell Cycle Regulation Antibody Sampler Kit II

Cell Cycle Regulation Antibody Sampler Kit II #9870

Western Blotting Image 1

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).

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Western Blotting 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.

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Western Blotting Image 3

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

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Western Blotting 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.

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Western Blotting Image 5

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.

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Western Blotting Image 6

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.

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Western Blotting Image 7

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

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Western Blotting Image 8

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).

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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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Western Blotting Image 10

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

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Western Blotting Image 11

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).

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Flow Cytometry Image 12

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.

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IHC-P (paraffin) Image 13

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

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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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IP Image 15

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.

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Flow Cytometry Image 16

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

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Flow Cytometry Image 17

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.

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IF-IC Image 18

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).

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IHC-P (paraffin) Image 19

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

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IP Image 20

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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IF-IC Image 21

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).

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IF-IC Image 22

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

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IHC-P (paraffin) Image 23

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

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IHC-P (paraffin) 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).

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IHC-P (paraffin) Image 25

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

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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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Flow Cytometry Image 27

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 28

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
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 

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.

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.

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.

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. Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.
  3. McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
  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.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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To Purchase # 9870T

Product Number Size Price
9870T 1 Kit (8 x 20 µl) $509.00.0
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