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Antibodies

RNA pol II antibody (mAb)

Clone: 4H8
Aliases: RPB1, RPO2, POLR2, POLRA, RPBh1, RPOL2, RpIILS, hsRPB1, hRPB220
Catalog No: 39097 Format: 200 µl $420 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG
Purification:Affinity Purified
Host:Mouse
Molecular Weight:217 kDa
Reactivity:Budding Yeast, C. elegans, Human, Mouse, Rat

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

for RNA pol II antibody (mAb) (Clone 4H8)Application Notes


Validated Applications:
ChIP: 10 µ1 per ChIP
ChIP-Seq: 20 µl each
WB: 1:2,000 - 1:5,000 dilution

Published Applications:
ChIP
ICC/IF
WB
See references for more information. Individual optimization may be required.

For optimal results in Western blotting, primary antibody incubations should be performed overnight at 4°C. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.

for RNA pol II antibody (mAb) (Clone 4H8)Immunogen

This RNA pol II antibody was raised against a synthetic peptide YSPTSpPS corresponding to human RNA pol II. It recognizes both the phosphorylated and unphosphorylated forms of the largest subunit of RNA pol II.

for RNA pol II antibody (mAb) (Clone 4H8)Buffer

PBS containing 0.2% gelatin, 0.01% thimersol and 0.1% sodium azide. Thimersol and sodium azide are highly toxic.

Also available as an AbFlex™ engineered recombinant antibody.  For details on the corresponding AbFlex Recombinant Antibody, see Catalog No. 91151.

for RNA pol II antibody (mAb) (Clone 4H8)References

 
RNA pol II antibody (mAb) tested by ChIP-Seq.

RNA pol II antibody (mAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 2.3 million HL-60 cells and 20 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 30 million sequence tags were mapped to identify Pol II binding. ChIP-Seq data from three specific genes is shown as an example. Pol II binding is detected at the 5´ end of the genes, throughout the gene bodies and at the 3´ end of the genes.

RNA pol II antibody (mAb) tested by Western blot.

RNA pol II antibody tested by Western blot.
RNA pol II detection by Western blot analysis. RNA pol II is detected in HeLa nuclear extracts using RNA pol II mouse monoclonal antibody at a 1:2,000 dilution.

for RNA pol II antibody (mAb) (Clone 4H8)Background

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.

for RNA pol II antibody (mAb) (Clone 4H8)Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for RNA pol II antibody.

for RNA pol II antibody (mAb) (Clone 4H8)qPCR Primers

Active Motif's Human Positive Control Primer Set ACTB-2 (Cat. No. 71005) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Positive Control Primer Set Actb-2 (Cat. No. 71017) and Mouse Negative Control Primer Set 1 (Cat. No. 71011) have been validated for both qPCR and endpoint PCR.

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 RNA pol II antibody (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use RNA pol II antibody (mAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with RNA pol II antibody (mAb).

for RNA pol II antibody (mAb) (Clone 4H8)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for RNA pol II antibody (mAb) (Clone 4H8)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

RNA pol II antibody (mAb)
 

Data Thumbnails

RNA pol II antibody (mAb) tested by ChIP-Seq.

ChIP-Seq of RNA pol II mAb.
(Click image to enlarge and see details.)

RNA pol II antibody (mAb) tested by Western blot.

Western blot of RNA pol II mAb.
(Click image to enlarge and see details.)