AbFlex RNA Pol II antibody (rAb)

Product Overview

• Article Name: AbFlex RNA Pol II antibody (rAb)
• Biozol Catalog Number: ACM-91151
• Supplier Catalog Number: 91151
• Alternative Catalog Number: ACM-91151
• Manufacturer: Active Motif
• Category: Antikörper

Additional Information

Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis Tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.

AbFlex® RNA Pol II antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.

RNA pol II (RNA polymerase II) is responsible for synthesizing messenger RNA in eukaryotes. RNA pol II 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, RNA pol II, in combination with several other polymerase subunits, form the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with AbFlex® RNA Pol II antibody (rAb).

 

Application Notes

Applications Validated by Active Motif:
CUT&Tag: 1 µl per 50 µl reaction
ChIP-Seq: 4 µg per ChIP
WB*: 0.1 – 1 µg/ml
IF: 2 µg/ml

*Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

AbFlex® recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental clone, see Catalog No. 39097.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP-Seq, ChIP-qPCR

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This RNA Pol II antibody was raised against a synthetic peptide YSPTSpPS corresponding to human RNA pol II. 

Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 35 mM NaOAc, 0.035% sodium azide, 30% glycerol.

 

ChIP-Seq of AbFlex® RNA Pol II antibody (rAb).
ChIP was performed using AbFlex RNA Pol II recombinant antibody with 20 µg chromatin from the HT1080 cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 12 million sequence tags were mapped to identify RNA Pol II binding sites. The image shows binding across a region of chromosome 12. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Western Blot of AbFlex® RNA Pol II antibody (rAb).
Western Blot: 30 µg HeLa cell nuclear extract* was run on an SDS-PAGE gel and probed with AbFlex RNA Pol II (rAb) at 1 µg/mL.
*Nuclear extract prepared with high salt / sonication protocol. Contact Technical Service for additional info.
MW: 217 kDa

Immunofluorescence stain of AbFlex® RNA Pol II antibody (rAb).
RNA Pol II was detected in human HeLa cells in a 2-step staining protocol using AbFlex RNA Pol II (rAb) at 2 µg/mL, followed by Anti-mouse IgG2a Dylight® 488.

 

Product Properties

• Clonality: Recombinant
• Tag: 6xHIS Tag, Avidin Tag

Images

ChIP-Seq of AbFlex® RNA Pol II antibody (rAb).

Western Blot of AbFlex® RNA Pol II antibody (rAb).

Immunofluorescence stain of AbFlex® RNA Pol II antibody (rAb).