Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
p15 INK4B Antibody 4822 40 µl 15 kDa Rabbit 
p27 Kip1 (SX53G8.5) Mouse mAb 3698 40 µl 27 kDa Mouse IgG1
p18 INK4C (DCS118) Mouse mAb 2896 40 µl 18 kDa Mouse IgG2a
p21 Waf1/Cip1 (12D1) Rabbit mAb 2947 40 µl 21 kDa Rabbit IgG
p57 Kip2 Antibody 2557 40 µl 57 kDa Rabbit 
p27 Kip1 (D69C12) XP® Rabbit mAb 3686 40 µl 27 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit provides an economical means to investigate cyclin dependent kinase inhibitors. The kit contains enough primary and secondary antibody to perform four western blot experiments.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

The cyclin-dependent kinase complex can exist as an active, binary complex containing cyclin and kinase or as an inactive, ternary complex when the pair associates with a CKI inhibitor. Some of these component proteins act selectively within their cyclin/CDK complex while other proteins exhibit a broader range of functions, including roles apart from the CDK complex (1,2). The INK4 family is characterized by 32 amino acid ankyrin repeats that mediate specific interactions with CDK4/6 and cyclinD/CDK4/6 (3). The Kip family includes p27, perhaps one of the most versatile proteins by virtue of its inherently context-adaptive structure. p27 has been implicated in regulatory roles of cell cycle progression, malignant transformation, cell motility, and differentiation (4,5). Similarly, p21 interacts with several CDK complexes during different cell cycle stages to exert different effects; p21 is regulated by phosphorylation and ubiquitin-mediated degradation (6,7).

  1. Harper, J.W. (1997) Cancer Surv 29, 91-107.
  2. Viallard, J.F. et al. (2001) Cancer Radiother 5, 109-29.
  3. Noh, S.J. et al. (1999) Cancer Res 59, 558-64.
  4. Borriello, A. et al. (2007) Cell Cycle 6, 1053-61.
  5. Chassot, A.A. et al. (2008) Cell Cycle 7, 2038-46.
  6. Kim, Y. et al. (2008) Genes Dev 22, 2507-19.
  7. Abbas, T. and Dutta, A. (2009) Nat Rev Cancer 9, 400-14.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #9867

    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit

    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from MCF-7 cells using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 2 Expand Image
    Flow cytometric analysis of Jurkat cells using p27 Kip1 (D69C12) XP® Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left) and Propidium Iodide (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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from HeLa cells, untreated or treated with dexamethasone (50 nM, 16h) alone or with λ phosphatase, using p57 Kip2 Antibody #2557.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from HeLa, 293 and Ramos cells, using p18 INK4C (DCS118) Mouse mAb.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 5 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 6 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis from control HeLa cells (lane 1) or HeLa cells with a targeted mutation in the gene encoding p27 Kip1 (lane 2) using p27 Kip1 (SX53G8.5) Mouse mAb (upper) or a-actinin (D6F6) XP Rabbit mAb #6487 (lower). The change in p27 Kip1 molecular weight in the mutated HeLa cells confirms the specificity of the antibody for p27 Kip1.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of whole cell extracts from 293, HeLa, HT29, C6, and NIH/3T3 cells, using p15 INK4B Antibody.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 9 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 10 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis, using a different p57 Kip2 mouse monoclonal antibody, of whole cell lysate from dexamethasone-treated (50 nM, 16h) HeLa cells (lane 1) and of p57 Kip2 immunoprecipitated from the same lysate using p57 Kip2 Antibody #2557 (lane 5). Lanes 2-4 represent controls for the immunoprecipitation experiment, as indicated.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from various cell types using p27 Kip1 (D69C12) XP® Rabbit mAb.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from NIH/3T3, C6 and MCF7 cells using p27 Kip1 (SX53G8.5) Mouse mAb.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 15 Expand Image
    Flow cytometric analysis of untreated C6 cells, using p15 INK4B Antibody versus propidium iodide (DNA content). Boxed population indicates p15 INK4B-positive cells.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 16 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 17 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 18 Expand Image
    Confocal immunofluorescent analysis of MCF7 cells using p27 Kip1 (SX53G8.5) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or dexamethasone-treated (right), using p57 Kip2 Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 20 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 21 Expand Image
    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).
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 22 Expand Image
    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).
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 23 Expand Image
    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.
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 24 Expand Image
    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).
    Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit: Image 25 Expand Image
    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.