Technical Data

R2039-21
Clone Type
Monoclonal
Host
Mouse
Isotype
IgG1
Clone Number
2Q2167 (ARNA-3)
Grade
Supernatant
Applications
IHC WB
Crossreactivity
Dr Hu
Accession #
NP_511124.1
Shipping Temp
Blue Ice
Storage Temp
-20°C
Mouse Anti-RNA Polymerase II (B220, DNA-directed RNA Polymerase II Largest Subunit, hRPB220, hsRPB1, POLR2, POLR2A, POLRA, RNA Polymerase II (DNA directed) Polypeptide A 220kD, RPB1, RPB220, RPBh1, RpIILS, RPO2, RPOL2)

DNA-directed RNA polymerase II subunit RPB1 (EC 2.7.7.6; UniProt P04052; also known as DNA-directed RNA polymerase III largest subunit, RNA polymerase II subunit B1) is encoded by the RpII215 (also known as CG1554) gene (Gene ID 32100) in drosophila melanogaster (fruit fly). RNA polymerase II (Pol II) is a multi-subunit enzyme responsible for the transcription of protein-coding genes. Transcription initiation requires recruitment of the complete transcription machinery to a promoter via solicitation by activators and chromatin remodeling factors. Pol II can coordinate 10 to 14 subunits. This complex interacts with the promoter regions of genes and a variety of elements and transcription factors. The DNA binding domain of the polymerase is a groove where TFIIB orients the DNA for unwinding and transcription.

Applications
Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested.
Recommended Dilution
Immunohistochemistry: 1:100; Frozen tissue; fixation recommendation - incubate tissue with 2% paraformaldehyde in PBS for 10min, wash in PBS for 5min, incubate in 0.2% Triton X-100 in PBS for 5min, wash in PBS for 5min. Incubate with antibody one hour. Optimal dilutions to be determined by the researcher.
Control
Positive for drosophila, kangaroo kidney epithelial cell line PtK2, and human HeLa and HEp-2 cell lines.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Purified Drosophila RNA polymerase II. Epitope: aa794-822
Form
Supplied as liquid, 0.09% sodium azide.
Purity
Supernatant
Specificity
Recognizes RNA Polymerase II.Excellent for detection of transcription activity, e.g. puffs in polytene chromosomes. Reacts with RNA polymerase II (Mr 175,000 subunit of RNAP II) in biological materials. Reactive polypeptides after SDS-PAGE: 200kD band of non-phosphorylated and 240kD band of phosphorylated Pol IIA. Reactivities on Cultured Cell Lines (tested so far): drosophila, kangaroo kidney epithelial cell line PtK2, and human HeLa and HEp-2 cell lines. Species Crossreactivity: kangaroo, human and drosophilia

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Dubois M-F, Bensaude O, et al. Heat shock of HeLa cells inactivates a nuclear protein phosphatase specific for dephosphorylation of the C-terminal domain of RNA polymerase II. Nucleic Acids Res 27: 1338-1344 (1999). |2. Nayler O, Stamm S, et al. SAF-B protein couples transcription and pre-mRNA splicing to SAR/MAR elements. Nucl Acids Res 26, 3542-3549 (1998).|3. Petersen G, Song D, Hügle-Dörr B, Oldenburg I and Bautz EKF: Mapping of linear epitopes recognized by monoclonal antibodies with gene-fragment phage display libraries. Mol. Gen. Genet. 249: 425-431.|4. Kontermann RE, Bautz EK, et al. (1995) Characterization of the epitope recognized by a monoclonal antibody directed against the largest subunit of Drosophila RNA polymerase II. Biol Chem Hoppe Seyler 376: 473-481.|5. Krämer, A., Bautz, E.K.F. et al. (1980) Monoclonal antibody directed against RNA polymerase II of drosophila melanogaster. Mol. Gen. Genet. 180: 193-199.
USBio References
No references available
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