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Applications Table
| Western Blot | Chromatin IP | IF | IP |
|---|---|---|---|
| 1:2,000 - 1:5,000 | 10 µl per ChIP | Yes | Yes |
for RNA pol II antibody (mAb)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)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)Application Notes
This RNA Polymerase II (RNA Pol II) antibody has been validated for use in chromatin IP (10 µl per ChIP) and Western blotting (1:2,000 - 1:5,000 dilution), and reported to work in ELISA, immunofluorescence and immunoprecipitation experiments. 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)Buffer
PBS containing 0.2% gelatin, 0.01% thimersol and 0.1% sodium azide. Thimersol and sodium azide are highly toxic.
for RNA pol II antibody (mAb)Positive Control
Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for RNA pol II antibody.
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
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).
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)Storage
Antibodies in solution can be stored at -80°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for RNA pol II antibody (mAb)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
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
