|
|
|
|
Western blot - PABP antibody (ab21060)
(enlarge)
|
|
Immunocytochemistry/ Immunofluorescence - PABP antibody (ab21060)
(enlarge)
|
|
Immunocytochemistry/ Immunofluorescence - PABP antibody (ab21060)
(enlarge)
|
|
|
|
|
Product Name
|
PABP antibody
|
| |
See all PABP antibodies (4)...
|
|
Product type
|
Primary antibodies
|
|
Description
|
Rabbit polyclonal to PABP
|
|
Immunogen
|
Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Human PABP.
(Peptide available as ab23710.)
|
|
Reacts with
|
Hu, Ms, Rat
|
|
Predicted to react
(species key)
|
Cow
(Due to sequence homology)
|
|
Tested applications
(see key)
|
ICC/IF, IHC-P, IP, WB
|
|
|
Abreviews
|
|
|
|
Application notes
(see key)
|
Recommended dilutions
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: 1/40. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP: Use at an assay dependent concentration.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 75 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
|
|
Positive control
(see definition)
|
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
|
|
Cellular localization
|
Cytoplasmic. Shuttles between the cytoplasm and the nucleus.
|
|
Research areas
|
Chromatin and Nuclear Signaling >> RNAi >> P bodies/GW bodies
Chromatin and Nuclear Signaling >> RNAi >> Dicer
Chromatin and Nuclear Signaling >> DNA / RNA >> Translation >> Regulation
|
|
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.
|
|
|
|
|
Database links
|
The links below go to external sites and will open in a new browser window |
|
Raised in
|
Rabbit
|
|
Clonality
|
Polyclonal
|
|
Isotype
|
IgG
|
|
Purity
|
Immunogen affinity purified
|
|
Storage buffer
|
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
|
|
Form
|
Liquid
|
|
Concentration
|
Batch dependent within range: 0.10 - 0.70 mg/ml
|
|
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.
|
| |
At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this PABP antibody is on this datasheet. But please do contact us if you would like any reassurance! |
 |
|
|
See below for PABP antibody images, references, products related to ab21060 and other tools.
|
| |
PABP antibody images:
|
 Western blot - PABP antibody (ab21060)
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
See Abreview
|
| |
Western blot - PABP antibody (ab21060)
All lanes : 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.
|
| |
- 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 PABP antibody (ab21060)
|
|
This product has been used in: (two most recent references)
Freibaum BD et al. Global analysis of TDP-43 interacting proteins reveals strong association with RNA splicing and translation machinery. J Proteome Res 9:1104-20 (2010). WB, ICC/IF. PubMed: 20020773
Paronetto MP et al. Sam68 regulates translation of target mRNAs in male germ cells, necessary for mouse spermatogenesis. J Cell Biol 185:235-49 (2009). WB; Mouse. PubMed: 19380878
See all 4 publication references for this product.
If you publish research using ab21060 please let us know so that we can cite the reference on this datasheet.
|
| |
|
Search PubMed (MEDLINE) for references to PABP
|
| |
PABP antibody - more information
|
Customer reviews (feedback) regarding PABP antibody |
Customer FAQs regarding PABP antibody |
Protocols for PABP antibody |
Price and availability of products related to PABP antibody |
| |
PABP antibody - related products:
|
|
|
| |
PABP antibody - other tools:
|
Contact Abcam with a Technical Enquiry about PABP antibody |
| |
All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Pricing information loading
