概述

  • 产品名称
  • 描述
    兔多克隆抗体to PABP
  • 经测试应用
    适用于: IP, Flow Cyt, WB, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Cow, Pig
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Human PABP.

    (Peptide available as ab23710.)

  • 阳性对照
    • 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

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 相关性
    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.
  • 细胞定位
    Cytoplasmic. Shuttles between the cytoplasm and the nucleus.
  • 数据库链接
  • 别名
    • 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 图像

  • 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.
  • 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).
  • Immunocytochemical analysis of rat OLN93 oligodendroglia cells, labeling PABP with ab21060. Cells were paraformaldehyde fized, permeabilized with 0.1% Triton X100, and blocked with 10% Normal Goat Serum for 1 hour at 20°C. Immonstaining with anti-PABP diluted 1/200 overnight at 4°C.

    See Abreview

  • 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.

     

  • 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

  • Immunocytochemical analysis of newborn porcine tracheal epithelial cells, labeling PABP with ab21060. Cells were formaldehyde fixed, premeabilized in cold methanol, and blocked with 5% horse serum in PBS for 40 minutes at 23°C. Immunostaining with primary anti-PABP diluted 1/250 for 12 hours at 4°C. 

    See Abreview

  • 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.or

Anti-PABP antibody (ab21060)参考文献

This product has been referenced in:
  • Wang H  et al. Embryonic lethal abnormal vision proteins and adenine and uridine-rich element mRNAs after global cerebral ischemia and reperfusion in the rat. J Cereb Blood Flow Metab 37:1494-1507 (2017). Rat . Read more (PubMed: 27381823) »
  • Arribas-Layton M  et al. The C-Terminal RGG Domain of Human Lsm4 Promotes Processing Body Formation Stimulated by Arginine Dimethylation. Mol Cell Biol 36:2226-35 (2016). WB ; Human . Read more (PubMed: 27247266) »

See all 31 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 May 17 2011

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