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Products:Epigenetics and Nuclear Signaling >> DNA / RNA >> Translation >> Regulation
Overview
Product name
Anti-PABP antibody
See all PABP products (4) ...
Description
Rabbit polyclonal to PABP
Tested applications
IP, WB, IHC-P, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Cow
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Human PABP.
(Peptide available as ab23710.)
Positive control
ab21060 gave a positive result in the following Human Whole Cell Lysates: Hela Mouse Whole Cell Lysates: NIH 3T3 MEF1 Mouse Tissue Lysates: Brain Pancreas Testis Spinal cord Ovary Rat Whole Cell Lysates: PC12 Rat Tissue Lysates: Brain
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> RNAi >> P bodies/GW bodies
Epigenetics and Nuclear Signaling >> RNAi >> Dicer
Epigenetics and Nuclear Signaling >> DNA / RNA >> Translation >> Regulation
Applications
Show applications key
Our Abpromise guarantee covers the use of ab21060 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IP: Use at an assay dependent concentration.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 75 kDa.
IHC-P: 1/40Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: Use a concentration of 1 µg/ml
Target
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.
Alternative names
- PAB 1 antibody
- PAB1 antibody
- PABP 1 antibody
see all
Database links
- Entrez Gene: 282296 Cow
- Entrez Gene: 26986 Human
- Entrez Gene: 18458 Mouse
- Entrez Gene: 171350 Rat
- Omim: 604679 Human
- SwissProt: P11940 Human
- SwissProt: P29341 Mouse
- SwissProt: Q9EPH8 Rat
see all
Anti-PABP antibody images:
Western blot - PABP antibody (ab21060)
Anti-PABP antibody (ab21060) at 1 µg/ml + HeLa whole cell lysate at 20 µg/ml
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 69 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)
ab21060 detects a band of ~ 75 kDa in HeLa whole cell lysate. A 75 kDa band is also seen in Jurkat and A431 whole cell lysates (data not shown). PABP is reported to have run at an apparent molecular mass of 73 kDa on SDS-PAGE. We believe the band we see at 75 kDa represents PABP.
Immunocytochemistry/ Immunofluorescence - PABP antibody (ab21060)
ICC/IF image of ab21060 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21060, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence. 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Immunocytochemistry/ Immunofluorescence - PABP antibody (ab21060)
ab21060 at 1/50 staining human HeLa cells by ICC/IF. HeLa cells fixed for 10 minutes with 1.9 % formaldehyde in 2 % sucrose in PBS then permeabilised for 10 minutes with 0.5% NP40 in PBS-10% sucrose. The cells were then blocked in horse serum and incubated with the antibody for 1 hour. An Alexa Fluor ® 555 conjugated donkey anti-rabbit antibody was used as the secondary.
This image is courtesy of an anonymous Abreview
Western blot - PABP antibody (ab21060)
All lanes : Anti-PABP antibody (ab21060) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lane 4 : Pancreas (Mouse) Tissue Lysate (ab29363)
Lane 5 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 6 : Spinal Cord (Mouse) Tissue Lysate
Lane 7 : Ovary (Mouse) Tissue Lysate - normal tissue
Lane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 9 : Brain (Rat) Tissue Lysate - normal tissue
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 69 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)
Additional bands at : 110 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PABP antibody (ab21060)
Image courtesy of Human Protein Atlas
ab21060 staining PABP in Human appendix. The paraffin embedded tissue was incubated with ab21060 (1/40 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab21060 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Further results for this antibody can be found at www.proteinatlas.org
References for Anti-PABP antibody (ab21060)
This product has been referenced in:
- Hasgall PAet al. The Putative RNA Helicase HELZ Promotes Cell Proliferation, Translation Initiation and Ribosomal Protein S6 Phosphorylation. PLoS One 6:e22107 (2011). WB; Human.Read more (PubMed: 21765940) »
- Damgaard CK & Lykke-Andersen J Translational coregulation of 5'TOP mRNAs by TIA-1 and TIAR. Genes Dev 25:2057-68 (2011).Read more (PubMed: 21979918) »
See all 11 publications for this product
Publishing research using ab21060? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"