WB, ChIP
H M Mk
Endogenous
200
Rabbit IgG
#O95602
25885
Product Information
Product Usage Information
For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Chromatin IP | 1:50 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro36 of human POLR1A protein.
Background
RNA polymerase I (RNAPI) is a large multi-protein complex that functions as a DNA dependent, RNA polymerase, which is primarily responsible for the transcription of ribosomal RNA (rRNA) genes. The largest subunit, Rpa194, or POLR1A, along with selectivity factor 1 (SL1), and the transactivator protein upstream binding factor (UBF) make up the core transcriptional machinery of the RNAPI complex (1-3). The RNAPI complex is recruited specifically to rDNA promoters by SL1, which contains TBP and TAF proteins, to transcribe precursors to rRNA (2). These precursors are processed into 18S, 5.8S, and 28S mature rRNAs, which make up most of the ribosomal structure (1). Similar to the RNA polymerase II complex, there are other core components that are required for transcription of target genes such as the TFIIH complex and SPT6 (4, 5). Overexpression of nascent rRNA has been shown to be associated with poor prognosis in certain cancer types, and enlarged nucleoli are a hallmark of cellular proliferation and aggressive tumors (6-8). Oncogenes such as Myc, Ras, and PI3K can drive RNAPI-mediated rRNA transcription, making POLR1A a key therapeutic target (9). Indeed, specific targeting of RNAPI activity with a small molecule inhibitor can induce autophagy selectively in tumor cells while having minimal effects in normal cells (10). Additionally, mutations in POLR1A are associated with acrofacial dysostosis, Cincinnati type, a cranioskeletal malformation syndrome. Loss of function mutations result in disrupted ribosome biogenesis and p53-mediated cell death affecting skeletal precursor cells or the neural-crest (11).
- Russell, J. and Zomerdijk, J.C. (2006) Biochem Soc Symp , 203-16.
- Jantzen, H.M. et al. (1990) Nature 344, 830-6.
- Comai, L. et al. (1992) Cell 68, 965-76.
- Iben, S. et al. (2002) Cell 109, 297-306.
- Engel, K.L. et al. (2015) Mol Cell Biol 35, 2321-31.
- Williamson, D. et al. (2006) Genes Chromosomes Cancer 45, 839-45.
- Derenzini, M. et al. (2000) J Pathol 191, 181-6.
- Maggi, L.B. and Weber, J.D. (2005) Cancer Invest 23, 599-608.
- Woods, S.J. et al. (2015) Biochim Biophys Acta 1849, 821-9.
- Drygin, D. et al. (2011) Cancer Res 71, 1418-30.
- Weaver, K.N. et al. (2015) Am J Hum Genet 96, 765-74.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting ChIP: Chromatin IP
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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