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Products:Epigenetics and Nuclear Signaling >> Transcription >> Polymerase associated factors >> Pol II Transcription >> Polymerase
Anti-PAF1 antibody - ChIP Grade (ab20662)
Product partner
Related products
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Proteins and Peptides
ab23389
ab23389
ab51501
ab52579
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Secondary antibodies
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DNA RNA and cDNA
ab46666
- Isotype controls
Reassurance, Refunds & Replacements
If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »Publishing research using ab20662? Please let us know so that we can cite the reference in this datasheet
ab20662 has been referenced in 3 publications.
- Wu J & Xu W Histone H3R17me2a mark recruits human RNA Polymerase-Associated Factor 1 Complex to activate transcription. Proc Natl Acad Sci U S A 109:5675-80 (2012). PubMed: 22451921
- Nakamura M et al. Spt6 levels are modulated by PAAF1 and proteasome to regulate the HIV-1 LTR. Retrovirology 9:13 (2012). WB; Human. PubMed: 22316138
- Ponnusamy MP et al. RNA polymerase II associated factor 1/PD2 maintains self-renewal by its interaction with Oct3/4 in mouse embryonic stem cells. Stem Cells 27:3001-11 (2009). CHIPseq; Mouse. PubMed: 19821493
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Western blot - PAF1 antibody (ab20662)
All lanes : Anti-PAF1 antibody - ChIP Grade (ab20662) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) (ab28446) at 1/10000 dilution
Predicted band size : 60 kDa
Observed band size : 85 kDa (why is the actual band size different from the predicted?)
Additional bands at : 73 kDa (possible cleavage fragment,cross reactivity).
ab20662 recognizes a band at approximately 80kDa, which corresponds in size to PAF1. Although it has a calculated molecular weight of ~60kDa, several groups have shown that it migrates at a size of 80-90kDa (see Moniaux, et.al., Oncogene Feb. 2006, Yart et.al., Mol Cell Biol 25:5052-60, 2005, and Rozenblatt-Rosen, et.al., Mol Cell Biol 25:612-20, 2005). There is also a smaller band at ~73kDa which may be a degradation product. Both bands are competed away by the addition of immunizing peptide, suggesting that the interaction is specific.
ChIP - PAF1 antibody (ab20662)
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 5µg of ab20662 (blue), and 20µl of Protein A/G sepharose beads in RIPA buffer. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH (active) and MYO-D and SERPINA (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
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