Immunohistochemistry analysis of RNA Polymerase II CTD showing staining in the nucleus and cytoplasm of paraffin-embedded human breast carcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a RNA Polymerase II CTD monoclonal antibody (Product # MA1-46093) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human, Mouse, Yeast|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Chemically synthesized phospho-ser 5 YSPTSpPS (Human).|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-10 ul|
|ELISA (ELISA)||1:100 - 1:2000|
|Immunocytochemistry (ICC)||1:50 - 1:100|
|Immunofluorescence (IF)||1:50 - 1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Immunoprecipitation (IP)||1:10 - 500|
|Western Blot (WB)||1:1000 - 1:20000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-46093 reacts with DNA-directed RNA Polymerase II Largest Subunit in human, yeast and mouse samples. Expected to cross-react with Drosophilia melanogaster, hamster, S. pombe, and Arabidopsis thaliana due to sequence homology.
MA1-46093 has been successfully used in Western Blot, Chromatin Immunoprecipitation, ELISA, Immunocytochemistry/Immunofluorescence, IHC (P) and Immunoprecipitation applications.
The MA1-46093 immunogen is 10 repeats of synthetic peptide YSPTSPS using chemically synthesized phospho-Ser 5 YSPTSpPS (Human). This antibody detects both the phosphorylated and non-phosphorylated forms.
The POLR2A gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. Recruitment of RNA processing factors to the elongation complex is coordinated by the phosphorylation of Serine-2 and Serine-5 residues within the C-terminal repeat (YSPTSPS) domain of the largest subunit of RNAPII. CTD4H8 can be used as a positive control in chromatin immunoprecipitation (ChIP) assays.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Hepatitis C virus infection activates an innate pathway involving IKK-¿ in lipogenesis and viral assembly.
MA1-46093 was used in ChIP assay to study the mechaism by which HCV infection activates IKK-alpha and stimulates lipogenesis and lipid droplet formation
|Li Q,Pène V,Krishnamurthy S,Cha H,Liang TJ||Nature medicine (19:722)||2013|
Lack of CAK complex accumulation at DNA damage sites in XP-B and XP-B/CS fibroblasts reveals differential regulation of CAK anchoring to core TFIIH by XPB and XPD helicases during nucleotide excision repair.
MA1-46093 was used in western blot to study the role of the XPB and XPD helicases in regulating the anchorage of the CAK complex to TFIIH during nucleotide excision repair
|Zhu Q,Wani G,Sharma N,Wani A||DNA repair (11:942)||2012|