Anti-PAF1/PD2 antibody - ChIP Grade (ab20662)
Key features and details
- Rabbit polyclonal to PAF1/PD2 - ChIP Grade
- Suitable for: WB, ChIP
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-PAF1/PD2 antibody - ChIP Grade
See all PAF1/PD2 primary antibodies -
Description
Rabbit polyclonal to PAF1/PD2 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: WB, ChIPmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Dog, Xenopus laevis
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Immunogen
Synthetic peptide corresponding to Human PAF1/PD2 aa 100-200 (internal sequence) conjugated to keyhole limpet haemocyanin.
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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.
- This antibody gave a positive signal in the following whole cell lysates: HeLa.
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General notes
Previously labelled as PAF1.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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
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Function
The mammalian PAF1 complex is a multifunctional complex. The PAF1 complex interacts with RNA polymerase II and may be involved in both initiation and elongation, and in histone methylation and RNA processing. Has oncogenic activity in vivo and in vitro. -
Sequence similarities
Belongs to the PAF1 family. -
Cellular localization
Nucleus. Punctuate distribution throughout the nucleus except in nucleoli and the perinuclear chromatin. - Information by UniProt
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Database links
- Entrez Gene: 54623 Human
- Entrez Gene: 54624 Mouse
- Entrez Gene: 361531 Rat
- Entrez Gene: 446278 Xenopus laevis
- Omim: 610506 Human
- SwissProt: Q8N7H5 Human
- SwissProt: Q8K2T8 Mouse
- SwissProt: Q4V886 Rat
see all -
Alternative names
- F23149 1 antibody
- FLJ11123 antibody
- hPAF1 antibody
see all
Images
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Western blot - Anti-PAF1/PD2 antibody - ChIP Grade (ab20662)All lanes : Anti-PAF1/PD2 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/PD2 peptide (ab23389) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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), 73 kDa (possible cross reactivity)ab20662 recognizes a band at approximately 80kDa, which corresponds in size to PAF1 / PD2. 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 - Anti-PAF1/PD2 antibody - ChIP Grade (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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
References (6)
ab20662 has been referenced in 6 publications.
- 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). 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 . PubMed: 26330467
- 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
- Ma MK et al. Histone crosstalk directed by H2B ubiquitination is required for chromatin boundary integrity. PLoS Genet 7:e1002175 (2011). PubMed: 21811414
- 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). ChIP-sequencing ; Mouse . PubMed: 19821493
Images
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Western blot - Anti-PAF1/PD2 antibody - ChIP Grade (ab20662)All lanes : Anti-PAF1/PD2 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/PD2 peptide (ab23389) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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), 73 kDa (possible cross reactivity)ab20662 recognizes a band at approximately 80kDa, which corresponds in size to PAF1 / PD2. 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 - Anti-PAF1/PD2 antibody - ChIP Grade (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.
