Invitrogen

CDK8 Polyclonal Antibody

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CDK8 Antibody in ChIP assay (ChIP) — Chromatin immunoprecipitation analysis of Cdk8 was performed using cross-linked chromatin from 1 x 106 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of a Cdk8 polyclonal antibody (Product # PA1-21780). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 of Egr1. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions); the zigzag line represents an intron; and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.

CDK8 Antibody — Figure 6. Transcriptional memory leads to Ssn3/Cdk8-dependent poised preinitiation complex. ( A ) ChIP against Med1-GFP, Med13-GFP or Ssn8-GFP from cells grown under repressing, activating or memory conditions. ( B ) ChIP against Ssn3-FRB-GFP from cells grown in repressing, activating or memory conditions. ( A and B ) *p<0.05, compared with the repressing condition (Student's t-test). ( C ) ChIP against RNAPII from strains expressing Ssn3-FRB-GFP grown under either repressing or memory conditions, before or after treatment 1 ug/ml of rapamycin using primers to amplify the INO1 promoter (-348 to -260) or the PRM1 CDS. Inset: confocal micrographs of Ssn3-FRB-GFP expressing cells before or after treatment with 1 mg/ml of rapamycin for 30 min. *p<0.05, compared with t = 0 (Student's t-test). ( D and E ) INO1 activation ( D ) or reactivation ( E ) in Ssn3-FRB-GFP cells. For activation at time = 0, cells were shifted from medium containing 100 muM inositol (repressing conditions; red arrow in schematic) to medium without inositol (activating conditions; green arrow in schematic). For reactivation, cells were shifted from activating medium to repressing medium containing 100 muM inositol for 3 hr. Cells were treated +-1 ug/ml rapamycin for 45 min before transferring to activating conditions. Cells were harvested at the indicated time points, and INO1 mRNA levels were quantified relative to ACT1 mRNA levels by RT-qPCR. The averages of three biological replicates are shown +- standard {TM}

产品细节

Applications

Tested Dilution

Publications

免疫印迹 (WB)

1-2 µg/mL

免疫细胞化学 (ICC/IF)

Assay-dependent

免疫沉淀 (IP)

10 µg/mg of lysate

染色质免疫沉淀实验 (ChIP)

1-3 µL

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产品规格

种属反应

Human, Mouse, Rat

已发表种属

Human

宿主/亚型

Rabbit / IgG

分类

Polyclonal

类型

Antibody

抗原

Synthetic peptide: C-SQQPPQYSHQTHRY, corresponding to amino acids 451-464 of Human Cdk8.

偶联物

Unconjugated

形式

Liquid

浓度

0.2 mg/mL

纯化类型

Protein A

保存液

10mM PBS, pH 7.4, with 0.2% BSA

内含物

0.09% sodium azide

保存条件

4° C, do not freeze

RRID

AB_2291488

产品详细信息

Recommended positive controls: HT29 cells.

靶标信息

A family of proteins designated as cdks are critical regulators of cell cycle progression. The prototype member of this family, p34/Cdc 2, and a related protein Cdk 2, function late in the cycle while Cdk 4 and Cdk 6 are critically involved in G1 to S progression. The p54/Cdk 8 interacts exclusively with Cyclin C. The Cdk 8 Cyclin C complex activates the kinase. Because of analogy with yeast Cdk 8, it has been proposed that the human Cdk 8 Cyclin C may interact with RNA polymerase, thus influencing the transcription mechanism(s) that lead to cell division.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

生物信息学

蛋白别名: CDK8 protein kinase; Cell division protein kinase 8; Cyclin-dependent kinase 8; Mediator complex subunit CDK8; Mediator of RNA polymerase II transcription subunit CDK8; MGC126074; MGC126075; Protein kinase K35

基因别名: CDK8; K35; RGD1560888

UniProt ID: (Human) P49336, (Mouse) Q8R3L8

Entrez Gene ID: (Human) 1024, (Mouse) 264064, (Rat) 498140

分子生物学功能: non-receptor serine/threonine protein kinase