Human DNA-directed RNA polymerase II subunit RPB1(POLR2A) ELISA kit

Unavailable
Code CSB-EL018327HU
See More Details 24T ELISA kits trial application
Product Type ELISA Kit
Size 96T,5×96T,10×96T
Uniprot No. P24928
Lead Time 5-7 working days
Abbreviation POLR2A
Protein Biological Process 1 Transcription/Transcription regulation
Target Name polymerase (RNA) II (DNA directed) polypeptide A, 220kDa
Alias MGC75453, POLR2, POLRA, RPB1, RPBh1, RPO2, RPOL2, RpIILS, hRPB220, hsRPB1, DNA-directed RNA polymerase II A|DNA-directed RNA polymerase II largest subunit, RNA polymerase II 220 kd subunit|DNA-direc
Species Homo sapiens (Human)
Protein Biological Process 3 Transcription
Sample Types serum, plasma, tissue homogenates, cell lysates
Detection Range 25 pg/mL-1600 pg/mL
Sensitivity 6.25 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Epigenetics and Nuclear Signaling
Protocol
Protocol may be improved. Please feel free to contact CUSABIO product specialist to obtain the latest version.
Assay Principle quantitative
Measurement Sandwich
Target Details This gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. The product of this gene contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, this subunit, in combination with several other polymerase subunits, forms the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA.
HGNC 9187
RGD 1587326
MGI 98086
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human POLR2A in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
SampleSerum(n=4)
1:1Average %95
Range %91-99
1:2Average %102
Range %97-107
1:4Average %87
Range %83-90
1:8Average %104
Range %101-109
Recovery
The recovery of human POLR2A spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample TypeAverage % RecoveryRange
Serum (n=5) 9589-98
EDTA plasma (n=4)8883-91
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/mlOD1OD2AverageCorrected
16002.331 2.398 2.365 2.199
8001.612 1.634 1.623 1.457
4000.978 0.968 0.973 0.807
2000.618 0.634 0.626 0.460
1000.419 0.432 0.426 0.260
500.299 0.278 0.289 0.123
250.235 0.216 0.226 0.060
00.167 0.165 0.166
References
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Function DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single-stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Regulation of gene expression levels depends on the balance between methylation and acetylation levels of tha CTD-lysines (By similarity). Initiation or early elongation steps of transcription of growth-factors-induced immediate early genes are regulated by the acetylation status of the CTD
Subcellular Location Nucleus, Cytoplasm
Protein Families RNA polymerase beta' chain family
Database Links

HGNC: 9187

OMIM: 180660

KEGG: hsa:5430

STRING: 9606.ENSP00000314949

UniGene: Hs.270017

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