Analysis Note

ControlPositive for drosophila, kangaroo kidney epithelial cell line PtK2, and human HeLa and HEp-2 cell lines.

Application

Western blot
Immunohistochemistry: 1:100; Frozen tissue; fixation recommendation - incubate tissue with 2% paraformaldehyde in PBS for 10 min, wash in PBS for 5 min, incubate in 0.2% Triton X-100 in PBS for 5 min, wash in PBS for 5 min. Incubate with antibody one hour.
Optimal working dilutions must be determined by the end user.

Detect RNA Polymerase II with Anti-RNA Polymerase II Antibody, clone ARNA-3 (Mouse Monoclonal Antibody), that has been shown to work in WB & IHC.

General description

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.

Immunogen

Purified Drosophila RNA polymerase II

Epitope: Amino acids 794-822

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

Supplied as hybridoma culture supernatant concentrate containing 0.09% sodium azide.

Specificity

Broad species cross-reactivity expected due to sequence homology.

ARNA-3 is excellent for detection of transcription activity, e.g. puffs in polytene chromosomes. Reacts with RNA polymerase II in biological materials. Reactive polypeptides after SDS-PAGE: 200 kDa band of nonphosphorylated and 240 kDa band of phosphorylated Pol IIA.