664908

antibody from BioLegend

Targeting: POLR2A POLR2, POLRA, RPB1

Immunocytochemistry (Data presented on provider website)

Immunoelectron microscopy (Supportive data in Antibodypedia)

Antibody Data

Product number

664908

Provider

BioLegend

Proper citation

BioLegend Cat#664908, RRID:AB_2566177

Product name

Alexa Fluor® 594 anti-RNA Polymerase II RPB1

Antibody type

Monoclonal

Antigen

Wheat germ RNA Polymerase II

Description

RPB1 is the catalytic and largest component of RNA polymerase II, which synthesizes mRNA precursors and many functional non-coding RNAs. It forms the polymerase active center together with RPB2, the second largest subunit. Polymerase II (Pol II) is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relatively to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft, and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single DNA template strand of the promoter is positioned within the central active site cleft of Pol II. Then, a bridging helix emanates from RPB1 and crosses the cleft near the catalytic site, which acts as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations during each neuocleotide addition. This promotes translocation of Pol II. Pol II moves on the template during transcription elongation. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II's largest subunit (RPB1), which serves as a platform for assembling factors that regulate transcription initiation, elongation, termination, and mRNA processing. It can act as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, being able to conform as both a replicate and transcriptase for the viral RNA circular genome.

Reactivity

Human, Mouse, Yeast

Host

Mouse

Conjugate

Yellow dye

Antibody clone number

8WG16

Vial size

100 µg

Concentration

0.5 mg/ml

Storage

2°C-8°C

Handling

Ambient RT

Immunoelectron microscopy

Supportive validation

Submitted by

BioLegend (provider)

Main image

Experimental details

F. HeLa cells were fixed with 2% paraformaldehyde (PFA) for ten minutes, permeabilized with 0.5% Triton X-100 for five minutes, and blocked with 5% FBS for 30 minutes. Then, the cells were intracellularly stained with 2 µg/ml Alexa Fluor® 594 conjugated (red) anti-RNA Polymerase II RPB1 (clone 8WG16 ) in a blocking buffer overnight at 4°C. Actin filaments were labeled with Alexa Fluor® 488 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.

Conjugate

Yellow dye