Anti-PABP antibody [10E10] (ab6125)
![Immunoprecipitation - Anti-PABP antibody [10E10] (ab6125)](https://www.abcam.com/ps/products/6/ab6125/Images/ab6125-12038-anti-pabp-antibody-10e10-immunoprecipitation.jpg "Immunoprecipitation - Anti-PABP antibody [10E10] (ab6125)")
Key features and details
- Mouse monoclonal [10E10] to PABP
- Suitable for: IP, ICC, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG2b
Overview
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Product name
Anti-PABP antibody [10E10]
See all PABP primary antibodies -
Description
Mouse monoclonal [10E10] to PABP -
Host species
Mouse -
Tested applications
Suitable for: IP, ICC, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Drosophila melanogaster -
Immunogen
Recombinant PABP (Human) expressed from its 1.85 kbp cDNA, NcoI to SspI.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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
Preservative: 0.1% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein A purified -
Purification notes
Purified from supernatant. -
Clonality
Monoclonal -
Clone number
10E10 -
Myeloma
Sp2/0 -
Isotype
IgG2b -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab6125 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 |
|---|---|---|
| IP |
Use at an assay dependent concentration.
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| ICC |
Use at an assay dependent concentration.
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| WB | (1) |
Use at an assay dependent concentration. Predicted molecular weight: 71 kDa.
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| Flow Cyt |
Use 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
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| Notes |
|---|
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IP
Use at an assay dependent concentration. |
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ICC
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 71 kDa. |
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Flow Cyt
Use 1µg for 106 cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
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Target
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Relevance
The poly(A)-binding protein (PABP), which is found complexed to the 3-prime poly(A) tail of eukaryotic mRNA, is required for poly(A) shortening and translation initiation. Grange et al. (1987) isolated a melanoma cell cDNA encoding human PABP. The predicted 633-amino acid protein contains 4 repeats of an approximately 80-amino acid unit in its N-terminal half. The authors found that this repeat region is highly conserved between human and yeast PABP and is sufficient for poly(A) binding. In vitro translation of the human PABP cDNA yielded a protein with an apparent molecular mass of 73 kD by SDS-PAGE. Northern blot analysis indicated that PABP is expressed as a 2.9-kb mRNA in human melanoma cells. Gorlach et al. (1994) noted that each of the 4 repeats of PABP is a ribonucleoprotein (RNP) consensus sequence RNA-binding domain. They determined that PABP has a pI of approximately 10.3 and is a very abundant, stable protein. Immunofluorescence studies of mammalian cells indicated that PABP is located exclusively in the cytoplasm. However, using both indirect immunofluorescence and tagging of PABP1 by fusion to the green fluorescent protein (GFP), Afonina et al. (1998) demonstrated that PABP1 shuttles between the nucleus and cytoplasm. PABP1 accumulated in the nucleus when transcription was inhibited, suggesting that active transcription is required for nuclear export of PABP1. -
Cellular localization
Cytoplasmic. Shuttles between the cytoplasm and the nucleus. -
Database links
- Entrez Gene: 26986 Human
- Omim: 604679 Human
- SwissProt: P11940 Human
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Alternative names
- PAB 1 antibody
- PAB1 antibody
- PABP 1 antibody
see all
Images
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Immunoprecipitation - Anti-PABP antibody [10E10] (ab6125)PABP was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to PABP (ab6125) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab6125.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 76kDa: PABP; 25kDa. -
![Immunocytochemistry - Anti-PABP antibody [10E10] (ab6125)](https://www.abcam.com/ps/products/6/ab6125/Images/ab6125-12037-anti-pabp-antibody-10e10-immunofluorescence.jpg)
Immunocytochemistry - Anti-PABP antibody [10E10] (ab6125)ICC/IF image of ab6125 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6125, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
![Flow Cytometry - Anti-PABP antibody [10E10] (ab6125)](https://www.abcam.com/ps/products/6/ab6125/Images/ab6125-12036-anti-pabp-antibody-10e10-flow-cytometry.jpg)
Flow Cytometry - Anti-PABP antibody [10E10] (ab6125)Overlay histogram showing HeLa cells stained with ab6125 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6125, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions. -
![Western blot - Anti-PABP antibody [10E10] (ab6125)](https://www.abcam.com/ps/products/6/ab6125/Images/ab6125-457448-anti-pabp-antibody-10e10-western-blot-hela-human.jpg)
Western blot - Anti-PABP antibody [10E10] (ab6125)Western Blot analysis on HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate using ab6125 at 1/100 to 1/6400 dilution. Anti-mouse IgG (whole molecule)-AP conjugate was used at the secondary at a 1/2000 dilution. Detection with BCIP/NBT substrate.
4-12% Bis-Tris, 1X MOPS running buffer.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (31)
ab6125 has been referenced in 31 publications.
- Li S et al. Chemosensitization of prostate cancer stem cells in mice by angiogenin and plexin-B2 inhibitors. Commun Biol 3:26 (2020). PubMed: 31942000
- Mateu-Regué À et al. Single mRNP Analysis Reveals that Small Cytoplasmic mRNP Granules Represent mRNA Singletons. Cell Rep 29:736-748.e4 (2019). PubMed: 31618640
- Yang X et al. Picornavirus 2A protease regulates stress granule formation to facilitate viral translation. PLoS Pathog 14:e1006901 (2018). PubMed: 29415027
- Haque N et al. ZFR coordinates crosstalk between RNA decay and transcription in innate immunity. Nat Commun 9:1145 (2018). PubMed: 29559679
- Xie X et al. Deubiquitylases USP5 and USP13 are recruited to and regulate heat-induced stress granules through their deubiquitylating activities. J Cell Sci 131:N/A (2018). PubMed: 29567855