The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Detects a band of approximately 270 kDa (predicted molecular weight: 217 kDa).
1/100 - 1/1000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration. PubMed: 22249179
Application notesIs unsuitable for IHC-P or IP.
RelevanceRPB1 is a subunit of RNA polymerase II which catalyses transcription of RNA from DNA. RPB1 forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative 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.
Overlay histogram showing HeLa cells stained with ab76123 (red line). The cells were fixed with 80% methanol (5 min)/ and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76123, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - RNA polymerase II antibody [EPR1509Y] (ab76123)
Anti-RNA polymerase II RPB1 antibody [EPR1509Y] - ChIP Grade (ab76123) at 1/2000 dilution + HeLa cell lysate at 10 µg
References for Anti-RNA polymerase II RPB1 antibody [EPR1509Y] - ChIP Grade (ab76123)
This product has been referenced in:
Kim JY et al. Chronic ethanol consumption inhibits glucokinase transcriptional activity by Atf3 and triggers metabolic syndrome in vivo. J Biol Chem289:27065-79 (2014).
Read more (PubMed: 25074928) »
Kim JW et al. New molecular bridge between RelA/p65 and NF-?B target genes via histone acetyltransferase TIP60 cofactor. J Biol Chem287:7780-91 (2012).
Read more (PubMed: 22249179) »