The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at 2-5 µg/mg of lysate.
Use at an assay dependent concentration.
Is unsuitable for WB.
Component of the Mediator complex, a coactivator involved in regulated gene transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. Phosphorylates the CTD (C-terminal domain) of the large subunit of RNA polymerase II (RNAp II), which may inhibit the formation of a transcription initiation complex. Phosphorylates CCNH leading to down-regulation of the TFIIH complex and transcriptional repression. Recruited through interaction with MAML1 to hyperphosphorylate the intracellular domain of NOTCH, leading to its degradation.
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily. Contains 1 protein kinase domain.
Immunoprecipitation of Cdk8 (3 µg of antibody per mg of HeLa cells lysate; 20% of the IP loaded). Lane 1- Immunoprecipitation with anti-Cdk8 antibody (ab115155); Lane 2-Immunoprecipitation with an anti-Cdk8 antibody that recognizes an upstream epitope; Lane 3- Immunoprecipitation with IgG control.
An anti-Cdk8 antibody used in lane 2 was used at 0.4 µg/ml for western blot. Detection: Chemiluminescence with exposure time of 30 sec.