Overview

  • Product nameAnti-PABP antibody
    See all PABP primary antibodies
  • Description
    Rabbit polyclonal to PABP
  • Tested applicationsSuitable for: IP, Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species 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

Applications

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.

Application Abreviews Notes
IP Use at an assay dependent concentration.
Flow Cyt 1/200. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa.
IHC-P 1/40. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • RelevanceThe 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 localizationCytoplasmic. Shuttles between the cytoplasm and the nucleus.
  • Database links
  • Alternative names
    • PAB 1 antibody
    • PAB1 antibody
    • PABP 1 antibody
    • PABP1 antibody
    • PABPC 1 antibody
    • PABPC1 antibody
    • PABPC2 antibody
    • PABPL1 antibody
    • Poly A binding protein 1 antibody
    • Poly A binding protein cytoplasmic 1 antibody
    • poly(A) binding protein, cytoplasmic 1 antibody
    • poly(A) binding protein, cytoplasmic 2 antibody
    • Polyadenylate binding protein 1 antibody
    see all

Anti-PABP antibody images

  • 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?)

    See Abreview

  • 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).
  • 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.

    See Abreview

  • All lanes : Anti-PABP antibody (ab21060) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate
    Lane 4 : Mouse pancreas tissue lysate - total protein (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.
  • 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:
  • Marno KM  et al. Novel restriction factor RNA-associated early-stage anti-viral factor (REAF) inhibits human and simian immunodeficiency viruses. Retrovirology 11:3 (2014). WB ; Human . Read more (PubMed: 24410916) »
  • Fathinajafabadi A  et al. CERKL, a retinal disease gene, encodes an mRNA-binding protein that localizes in compact and untranslated mRNPs associated with microtubules. PLoS One 9:e87898 (2014). WB, ICC/IF ; Human . Read more (PubMed: 24498393) »

See all 24 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Pig Cell (Newborn Porcine Tracheal Epithelial cell line)
Permeabilization Yes - Cold methanol
Specification Newborn Porcine Tracheal Epithelial cell line
Blocking step Serum as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative Formaldehyde
Username

Sathya Thulasi Raman

Verified customer

Submitted Jan 11 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Fibroblast)
Permeabilization No
Specification Fibroblast
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Methanol
Username

Abcam user community

Verified customer

Submitted Jun 22 2015

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (cg4 cell line)
Specification cg4 cell line
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jan 03 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample Rat Cell (OLN93 oligodendroglia cell line)
Specification OLN93 oligodendroglia cell line
Permeabilization Yes - 0,1% Triton X100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (HeLa)
Specification HeLa
Preparation Cell harvesting/tissue preparation method: grown in TC
Sample buffer: harvested by scraping in cold PBS
Fixation BD CytoFix/CytoPerm
Permeabilization Yes - BD CytoFix/CytoPerm
Gating Strategy single cells
Username

Lee Gehrke

Verified customer

Submitted Apr 10 2013

Thank you for your inquiry.

The peptide ab23710 is only 13 a.a. long. The listing of the "Protein length" on the datasheet is how long the full length protein is according to Swiss Prot (636 a.a. long).

http://www.uniprot....

Read More
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application CLIP
Sample Human Cell lysate - whole cell (HEK293T)
Specification HEK293T
Type iCLIP
Irradiation Energy Level (mJ/cm2): 150
Wavelength (nm): 254
Positive control Anti-hnRNP C antibody (sc-15386) bound protein A dynabeads, 0.1 mg/ml (antibody/beads)
Negative control Protein A dynabeads without antibody bound
Immuno-precipitation step Other - Protein A dynabeads
Username

Abcam user community

Verified customer

Submitted Mar 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLA)
Loading amount 60 µg
Specification HeLA
Gel Running Conditions Reduced Denaturing (4-15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jul 27 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblasts)
Loading amount 25 µg
Specification Mouse Embryonic Fibroblasts
Gel Running Conditions Reduced Denaturing (4-15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jul 27 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (mouse adipocyte)
Loading amount 5 µg
Specification mouse adipocyte
Gel Running Conditions Reduced Denaturing (8% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 17 2011

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"