Overview

  • Product nameAnti-PAF1 antibody - ChIP Grade
    See all PAF1 primary antibodies
  • Description
    Rabbit polyclonal to PAF1 - ChIP Grade
  • Tested applicationsSuitable for: WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Dog, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human PAF1.

    (Peptide available as ab23389.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa.

      This antibody gave a positive signal in chromatin prepared from the following whole cell lysates: HeLa.

Properties

Applications

Our Abpromise guarantee covers the use of ab20662 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 60 kDa).
ChIP Use 5-25 µg for µg of chromatin.

Target

Anti-PAF1 antibody - ChIP Grade images

  • 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 Human PAF1 peptide (ab23389) at 1 µg/ml

    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.
  • 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. 

References for Anti-PAF1 antibody - ChIP Grade (ab20662)

This product has been referenced in:
  • 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). Read more (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 . Read more (PubMed: 22316138) »

See all 4 Publications for this product

Product Wall


We have these 3 forward and reverse primers that were used for validation, in our catalog:

promoter region: ab83623, ab83624
intron 3: ab83625, ab83626
exon 6: ab83627, ab83628

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"