製品の概要

  • 製品名
    Anti-PAF1 antibody - ChIP Grade
    PAF1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to PAF1 - ChIP Grade
  • アプリケーション
    適用あり: WB, ChIPmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: 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.)

  • ポジティブ・コントロール
    • 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

画像

  • 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. 

参考文献

This product has been referenced in:
  • Cao QF  et al. Characterization of the Human Transcription Elongation Factor Rtf1: Evidence for Nonoverlapping Functions of Rtf1 and the Paf1 Complex. Mol Cell Biol 35:3459-70 (2015). Read more (PubMed: 26217014) »
  • Yao X  et al. The Mediator subunit MED23 couples H2B mono-ubiquitination to transcriptional control and cell fate determination. EMBO J 34:2885-902 (2015). WB, ChIP . Read more (PubMed: 26330467) »

See all 6 Publications for this product

レビューと Q&A


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"

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