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PDP - Template Name: Antibody Sampler Kit
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Cell Cycle/Checkpoint Antibody Sampler Kit #9917

Cell Cycle/Checkpoint Antibody Sampler Kit: Image 1
Western blot analysis of extracts from HeLa cells, untreated or UV-treated, using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

To Purchase # 9917

Cat. # Size Qty. Price Ships
9917T 1 Kit
6 x 20 microliters
$558
  • Product Includes
  • Related Products
Product IncludesQuantityApplicationsReactivityMW(kDa)Isotype
Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb #453920 µlWB, IP, IF, FH M R Mk34Rabbit 
Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #234820 µlWB, IF, FH M R Mk56Rabbit IgG
Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb #219720 µlWB, IP, IHC, FH62Rabbit IgG
Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb #851620 µlWB, IP, IHC, IF, FH M R Mk110Rabbit IgG
Phospho-Rb (Ser795) Antibody #930120 µlWB, IPH Mk110Rabbit 
Phospho-p53 (Ser15) (16G8) Mouse mAb #928620 µlWB, IF, FH53Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody #7074100 µlWBRabGoat 
Anti-mouse IgG, HRP-linked Antibody #7076100 µlWBMHorse 

Product Information

Product Description

The Cell Cycle/Checkpoint Antibody Sampler Kit provides a fast and economical means of evaluating multiple proteins involved in the cell cyle and checkpoint control. The kit contains enough primary and secondary antibody to perform four Western blot experiments.

Specificity / Sensitivity

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. Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb detects endogenous levels of Chk2 only when phosphorylated at Thr68. Phospho-Chk1 (Ser345) Antibody detects Chk1 only when phosphorylated at Ser345 and does not cross-react with other proteins. Phospho-Rb (Ser795) Antibody detects Rb only when phosphorylated at Ser795 and does not cross-react with Rb phosphorylated at other sites. Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb recognizes endogenous levels of Rb protein only when phosphorylated at Ser807, Ser811, or at both sites. This antibody does not cross-react with Rb phosphorylated at Ser608. Phospho-p53 (Ser15) (16G8) Mouse mAb detects endogenous levels of p53 only when phosphorylated at Ser15. The antibody does not cross-react with p53 phosphorylated at other sites.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser795 of human Rb. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser807/811 of human Rb protein, residues surrounding Ser345 of human Chk1, residues surrounding Ser15 of human p53, residues surrounding Tyr15 of human cdc2, and residues surrounding Thr68 of human Chk2.

Background

The cell division cycle demands accuracy to avoid the accumulation of genetic damage. This process is controlled by molecular circuits called "checkpoints" that are common to all eukaryotic cells (1). Checkpoints monitor DNA integrity and cell growth prior to replication and division at the G1/S and G2/M transitions, respectively. The cdc2-cyclin B kinase is pivotal in regulating the G2/M transition (2,3). Cdc2 is phosphorylated at Thr14 and Tyr15 during G2-phase by the kinases Wee1 and Myt1, rendering it inactive. The tumor suppressor protein retinoblastoma (Rb) controls progression through the late G1 restriction point (R) and is a major regulator of the G1/S transition (4). During early and mid G1-phase, Rb binds to and represses the transcription factor E2F (5). The phosphorylation of Rb late in G1-phase by CDKs induces Rb to dissociate from E2F, permitting the transcription of S-phase-promoting genes. In vitro, Rb can be phosphorylated at multiple sites by cdc2, cdk2, and cdk4/6 (6-8). DNA damage triggers both the G2/M and the G1/S checkpoints. DNA damage activates the DNA-PK/ATM/ATR kinases, which phosphorylate Chk at Ser345 (9), Chk2 at Thr68 (10) and p53 (11). The Chk kinases inactivate cdc25 via phosphorylation at Ser216, blocking the activation of cdc2.

Pathways

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