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9867
Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Cyclin Dependent Kinase Inhibitor Antibody Sampler Kit #9867

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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.
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.
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.
Western blot analysis of extracts from HeLa, 293 and Ramos cells, using p18 INK4C (DCS118) Mouse mAb.
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.
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.
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.
Western blot analysis of whole cell extracts from 293, HeLa, HT29, C6, and NIH/3T3 cells, using p15 INK4B Antibody.
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.
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.
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.
Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb.
Western blot analysis of extracts from various cell types using p27 Kip1 (D69C12) XP® Rabbit mAb.
Western blot analysis of extracts from NIH/3T3, C6 and MCF7 cells using p27 Kip1 (SX53G8.5) Mouse mAb.
Flow cytometric analysis of untreated C6 cells, using p15 INK4B Antibody versus propidium iodide (DNA content). Boxed population indicates p15 INK4B-positive cells.
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.
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.
Confocal immunofluorescent analysis of MCF7 cells using p27 Kip1 (SX53G8.5) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
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).
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.
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).
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).
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.
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).
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.
Inquiry Info.# 9867

Product 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.

Specificity / Sensitivity

Each antibody in the Cyclin Dependent Kinase Inhibitor Sampler Kit detects endogenous levels of its respective target protein and does not cross-react with other family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues within the amino terminus of p15, the carboxy terminus of human p16 or the carboxy terminus of p57. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues within the carboxy terminus of human p21, the amino terminus of p27 (#3686), and recombinant p27 (#3698).

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).

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For Research Use Only. Not for Use in Diagnostic Procedures.
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