Anti-PAF1抗体- ChIP Grade (ab20662)

概述

  • 产品名称Anti-PAF1抗体- ChIP Grade
    参阅全部 PAF1 一抗
  • 描述
    兔多克隆抗体to PAF1 - ChIP Grade
  • 经测试应用适用于: WB, ChIPmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat, Dog, Xenopus laevis
  • 免疫原

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

    (Peptide available as ab23389.)

  • 阳性对照
    • 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.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

Anti-PAF1 antibody - ChIP Grade 图像

  • 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. 

Anti-PAF1 antibody - ChIP Grade (ab20662)参考文献

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"