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Western blot - PAF1 antibody (ab20662)
(enlarge)
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ChIP - PAF1 antibody (ab20662)
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Product Name
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PAF1 antibody - ChIP Grade
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See all PAF1 antibodies (5)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to PAF1 - ChIP Grade
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human PAF1.
(Peptide available as ab23389.)
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Reacts with
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Hu
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Predicted to react
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Ms, Rat, Dog, Xl
(Due to sequence homology)
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Tested applications
(see key)
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ChIP, WB
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Abreviews
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Application notes
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Recommended dilutions
ChIP: Use 5µg for every 25µg of chromatin.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately ~80 kDa (predicted molecular weight: 60 kDa). Can be blocked with PAF1 peptide (ab23389).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Chromatin and Nuclear Signaling >> Transcription >> Elongation factors
Chromatin and Nuclear Signaling >> Transcription >> Cancer susceptibility >> Proto-oncogenes
Chromatin and Nuclear Signaling >> Transcription >> Polymerase associated factors >> Pol II Transcription >> Polymerase
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Relevance
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In yeast, PAF1 acts as a docking protein and is involved in transcriptional elongation through interactions with RNA polymerase II and other members of the transcription machinery. The human homolog has also been shown to interact with RNA polymerase II, in particular with the form which is phosphorylated on Serine 2 or 5 in the carboxy-terminal domain (CTD). It also associates with HRPT2 (parafibromin) and other members of the PAF1 complex (LEO1 and CTR9). This complex is involved in transcriptional elongation and 3' end processing. There is evidence that overexpression of PAF1 is involved in cancer development.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.40 - 0.50 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this PAF1 antibody - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for PAF1 antibody - ChIP Grade images, references, products related to ab20662 and other tools.
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PAF1 antibody - ChIP Grade images:
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 Western blot - PAF1 antibody (ab20662)
All lanes : 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 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.
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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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Search PubMed (MEDLINE) for references to PAF1
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PAF1 antibody - ChIP Grade - more information
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Customer reviews (feedback) regarding PAF1 antibody - ChIP Grade |
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PAF1 antibody - ChIP Grade - related products:
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PAF1 antibody - ChIP Grade - other tools:
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