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Chromatin immunoprecipitation analysis of Casein Kinase II alpha 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 1.0ul/100ul well volume of a Casein Kinase II alpha monoclonal antibody (Product # LF-MA0223). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1ul of eluted DNA in 2ul 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.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG2a, kappa|
|Immunogen||Recombinant human protein purified from E.coli(His-Casein kinase II a)|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
A suggested positive control for this product is Jurkat T cells.
Casein kinase II is a serine/threonine protein kinase that phosphorylates acidic proteins such as casein. The kinase exists as a tetramer and is composed of an alpha, an alpha-prime, and two beta subunits. The alpha subunits contain the catalytic activity while the beta subunits undergo autophosphorylation. The protein encoded by this gene represents the alpha subunit. While this gene is found on chromosome 20, a related transcribed pseudogene is found on chromosome 11. Three transcript variants encoding two different proteins have been found for this gene.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
casein kinase 2, alpha 1 polypeptide; casein kinase II alpha 1 polypeptide pseudogene; casein kinase II alpha 1 subunit; casein kinase II subunit alpha; casein kinase II, alpha 1 polypeptide; casein kinase II, alpha 1 related sequence 4; CK II alpha; CK II alpha 3; CK2 catalytic subunit alpha; CK2A1; CKII; protein kinase CK2
CK2A1; CKII; Ckiia; CSNK2A1; Csnk2a1-rs4; CSNK2A3