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RNA pol II - going, going, paused...
RNA-polymerase II (Pol II)-mediated transcription is a highly regulated process that determines cellular function and cell identity through the synthesis of mRNAs. RNA Pol II regulation at the stage of initiation is a key mechanism to control gene expression, so that a thorough understanding of gene regulation includes the actual process of transcription and this key driver of the process - RNA Pol II initiation, pausing, and active elongation.
Active Motif offers a unique suite of antibodies designed to identify RNA Pol II, and those C-terminal domains (CTDs) whose phosphorylation states are critical for enzyme activity. Another important approach is to measure RNA Pol II occupancy across the genome using ChIP, which provides information on start site selection, pre-initiation complex (PIC) formation, pausing, and transcription rates. Our TranscriptionPath™ Assay is uniquely suited for this type of analysis.
Antibodies List
| Product | Application | Format | Cat. No. |
|---|---|---|---|
| AbFlex® RNA Pol II antibody (rAb) | ChIP-Seq, IF, WB | 10 µg | 91152 |
| 100 µg | 91151 | ||
| RNA pol II antibody (mAb) | ChIP, ChIP-Seq, ICC, IF, WB | 50 µl | 39497 |
| 200 µl | 39097 | ||
| RNA pol II antibody (mAb) | WB | 10 µg | 61082 |
| 100 µg | 61081 | ||
| RNA pol II antibody (mAb) | ChIP, ChIP-Seq, ICC, IF, WB | 10 µg | 61668 |
| 100 µg | 61667 | ||
| AbFlex® RNA Pol II CTD phospho Tyr1 antibody (rAb) | ELISA, IF, WB | 10 µg | 91220 |
| 100 µg | 91219 | ||
| RNA Pol II CTD phospho Tyr1 antibody (mAb) | ChIP, IP, WB | 10 µg | 61384 |
| 100 µg | 61383 | ||
| AbFlex® RNA pol II CTD phospho Ser2 antibody (rAb) | ChIP, ChIP-Seq, ELISA, ICC, IF, WB | 10 µg | 91116 |
| 100 µg | 91115 | ||
| RNA pol II CTD phospho Ser2 antibody (mAb) | ChIP, ChIP-Seq, ICC, IF, IP, WB | 10 µg | 61084 |
| 100 µg | 61083 | ||
| RNA pol II CTD phospho Ser2 antibody (mAb) | ICC, IF, WB | 10 µg | 61700 |
| 100 µg | 61699 | ||
| RNA pol II CTD phospho Ser2 antibody (pAb) | IP, WB | 10 µl | 39564 |
| 200 µl | 39563 | ||
| RNA pol II CTD Ser2ph / Ser5ph antibody (mAb) | ChIP, ChIP-Seq, ICC, IF, WB | 10 µg | 61666 |
| 100 µg | 61665 | ||
| RNA Pol II CTD phospho Thr4 antibody (mAb) | ChIP, DB, ICC, IF, IP, WB | 10 µg | 61362 |
| 100 µg | 61361 | ||
| RNA Pol II CTD phospho Thr4 antibody (pAb) | WB | 10 µg | 61308 |
| 100 µg | 61307 | ||
| AbFlex® RNA pol II CTD phospho Ser5 antibody (rAb) | ChIP, ChIP-Seq, ELISA, ICC, IF, WB | 10 µg | 91120 |
| 100 µg | 91119 | ||
| RNA pol II CTD phospho Ser5 antibody (mAb) | ChIP, ChIP-Seq, ICC, IF, IP, WB | 10 µg | 61086 |
| 100 µ | 61085 | ||
| RNA pol II CTD phospho Ser5 antibody (mAb) | ICC, IF, WB | 10 µg | 61702 |
| 100 µg | 61701 | ||
| RNA pol II CTD phospho Ser5 antibody (pAb) | ChIP, ICC, IF, WB | 10 µl | 39234 |
| 100 µl | 39233 | ||
| RNA pol II CTD phospho Ser5 antibody (pAb) | ChIP, ChIP-Seq, WB | 10 µg | 39750 |
| 100 µg | 39749 | ||
| RNA pol II CTD phospho Ser7 antibody (mAb) | ChIP, ChIP-Seq, ICC, IF, IP, WB | 10 µg | 61088 |
| 100 µg | 61087 | ||
| RNA pol II CTD phospho Ser7 antibody (mAb) | ICC, IF, WB | 10 µg | 61704 |
| 100 µg | 61703 |
TranscriptionPath™
TranscriptionPath™ assay provides an alternate approach to RNA-based gene expression methods. TranscriptionPath is a Chromatin Immunoprecipitation-based assay used to measure transcription rates as a function of RNA Polymerase II occupancy across the genome. Unlike other RNA-based techniques, the method enables you to measure transcription rates without the influence of RNA half-life, and with the added advantage of using the same material that is used in your transcription factor targeted ChIP experiments. So now you can correlate transcription factor binding with transcriptional changes in the same experiment.
- Measure transcription rates without the influence of RNA half-life.
- Ideal for measuring changes in gene expression at early time points (minutes).
- Detect alternate start sites and unannotated genes.
- Identify genes that are poised for transcriptional activation.
- Perform in parallel with transcription factor-targeted ChIP to correlate TF binding with changes in transcription.
TranscriptionPath data reveals four states of RNAPII occupancy.
TranscriptionPath-Seq was performed using chromatin from mouse neuroblastoma cells. Four patterns of RNAPII occupancy were detected; examples are shown above. (A) Promoter Paused and Transcribed -- Transcription is rate-limited post-initiation. (B) Not Paused and Transcribed -- Transcription is rate-limted by RNAAPII recruitment. (C) Promoter Paused and Not Transcibed - These genes are poised for activation, but are not transcribed. In this example, the posed pattern shown is inducible. (D) Not Paused and Not Transcribed - These genese are inactive.
