Overview

  • Product name
    Anti-PTBP1 antibody [SH54]
    See all PTBP1 primary antibodies
  • Description
    Mouse monoclonal [SH54] to PTBP1
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    HeLa nuclear extract (Human)

  • Positive control
    • IHC-P: human breast carcinoma WB: Jurkat cell lysate, HeLa nuclear lysate ICC: Jurkat, HeLa cells

Properties

Applications

Our Abpromise guarantee covers the use of ab30317 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
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 57 kDa.
IHC-P Use a concentration of 2.5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 - 3 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Plays a role in pre-mRNA splicing and in the regulation of alternative splicing events. Binds to the polypyrimidine tract of introns. May promote RNA looping when bound to two separate polypyrimidine tracts in the same pre-mRNA. May promote the binding of U2 snRNP to pre-mRNA. Cooperates with RAVER1 to modulate switching between mutually exclusive exons during maturation of the TPM1 pre-mRNA.
  • Sequence similarities
    Contains 4 RRM (RNA recognition motif) domains.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 57 kDa RNA binding protein PPTB 1 antibody
    • 57 kDa RNA-binding protein PPTB-1 antibody
    • Heterogeneous nuclear ribonucleoprotein I antibody
    • Heterogeneous Nuclear Ribonucleoprotein Polypeptide I antibody
    • hnRNP I antibody
    • HNRNP-I antibody
    • HNRNPI antibody
    • HNRPI antibody
    • MGC10830 antibody
    • MGC8461 antibody
    • Polypyrimidine tract binding protein (heterogeneous nuclear ribonucleoprotein I) antibody
    • Polypyrimidine Tract Binding Protein 1 antibody
    • Polypyrimidine tract binding protein antibody
    • Polypyrimidine tract-binding protein 1 antibody
    • pPTB antibody
    • PTB 1 antibody
    • PTB 2 antibody
    • PTB 3 antibody
    • PTB 4 antibody
    • PTB antibody
    • PTB T antibody
    • PTB1 antibody
    • PTB2 antibody
    • PTB3 antibody
    • PTB4 antibody
    • PTBP 1 antibody
    • PTBP1 antibody
    • PTBP1_HUMAN antibody
    • PTBT antibody
    • RNA Binding Protein antibody
    see all

Images

  • Overlay histogram showing HeLa cells stained with ab30317 (red line). The cells were fixed with 80% 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. The cells were then incubated with the antibody (ab30317, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 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.

References

This product has been referenced in:
  • Gao W  et al. La Autoantigen Induces Ribosome Binding Protein 1 (RRBP1) Expression through Internal Ribosome Entry Site (IRES)-Mediated Translation during Cellular Stress Condition. Int J Mol Sci 17:N/A (2016). Read more (PubMed: 27447629) »
  • Cosme RS  et al. Roles of polypyrimidine tract binding proteins in major immediate-early gene expression and viral replication of human cytomegalovirus. J Virol 83:2839-50 (2009). WB, ICC/IF ; Human . Read more (PubMed: 19144709) »
See all 4 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your enquiry. The antibody was raised in mice immunized with HeLa nuclear protein extract as described and shown in this publication: http://www.jcb.org/cgi/content/full/137/5/965 It's a protein A purified mouse monoclonal IgG1. This clone was never specifically epitope mapped so, if there's any sequence homology between isoforms 1 and 2 and this IgG detects any of the epitopes they have in common, then cross-reactivity would not be an unexpected result. There have been some publications using this clone and you can refer to them should you require more information. Huang S et al. The perinucleolar compartment and transcription. J Cell Biol 143:35-47 (1998). PubMed: 9763419 Huang S et al. The dynamic organization of the perinucleolar compartment in the cell nucleus. J Cell Biol 137:965-74 (1997). PubMed: 9166399

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Lymphoblast cell line GM10725)
Specification
Human Lymphoblast cell line GM10725
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5%

Dr. Zeeshan Ali

Verified customer

Submitted Dec 06 2007

Answer

I'm sorry to hear you are experiencing problems with ab30317. I have been in contact with the laboratory that tested the antibody and received information which I hope will be useful: PTB expression can be high and at some point during growth and development all cells will need to express PTB so that in one sense, it can be said to be ubiquitously expressed. Perinucleolar compartments (PNC) - where PTB is localized - is predominantly present in cancer cells and rarely present in normal cells. The PNC, if present in normal cells, is much smaller and has a lower expression of PTB than the PNC of cancer cells. Therefore, if you are working with normal human tissues (the antibody does not detect rat or mouse protein), it is possible that PTB levels will be low, perhaps undetectable. Another factor to consider is tissue handling - some proteins are degraded more rapidly than others, especially those regulated in a cell cycle-dependent manner. I would like to recommend to use the positive control of human breast carcinoma sections. In the laboratory the antibody was used at a concentration of 2.5 ug/ml using DAB detection system and included a heat pre-treatment step for antigen retrieval using citrate buffer. Antigen retrieval can damage the epitope if it is too long, I would therefore recommend to try a shorter period. As the protein is located in the nucleus I would like to recommend to add tritonX100 (0.3%v/v) to your blocking buffer and antibody dilution buffer (TBS or PBS, though we prefer PBS for IHC experiments) to promote the permeabilisation of the antibody into the cells. Finally, it may be worth trying a traditional ABC amplification method rather than the Impress kit, as we have no experience of this kit and it may be not suitable for this antibody. I hope the above recommendations will help you, please do not hesitate to contact me again if you still experience problems with the positive control, I would be happy to offer you a replacement vial or refund if the antibody was purchased in the last 90 days,

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

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