Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PTBP1 antibody [EPR9048(B)] (ab133734)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9048(B)] to PTBP1
  • Suitable for: ChIP, WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PTBP1 antibody [EPR9048(B)]
    See all PTBP1 primary antibodies

  • Description

    Rabbit monoclonal [EPR9048(B)] to PTBP1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP, WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide corresponding to a region within Human PTBP1 (UniProt ID: P26599).

  • Positive control

    • WB: HeLa, HAP1, Jurkat, Daudi, A549 and BxPC3 cell lysates. Human fetal liver tissue lysate. IHC-P: Human breast carcinoma tissue. Flow Cyt: A549 cells.

  • General notes

     

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab133734 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
ChIP Use at an assay dependent concentration. PubMed: 27252533
WB 1/10000 - 1/50000. Predicted molecular weight: 57 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Perform antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/2000.
ICC/IF 1/250 - 1/500.
  • Application notes
    Is unsuitable for IP.

    • 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

      see all

    • 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

    • Western blot - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Western blot - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      All lanes : Anti-PTBP1 antibody [EPR9048(B)] (ab133734) at 1/10000 dilution

      Lane 1 : Wild-type HeLa lysate
      Lane 2 : PTBP1 knockout HeLa lysate
      Lane 3 : Wild-type HAP1 lysate
      Lane 4 : PTBP1 knockout HAP1 lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 57 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab133734 observed at 57 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab133734 Anti-PTBP1 antibody [EPR9048(B)] was shown to specifically react with PTBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265155 (knockout cell lysate ab257614) was used. Wild-type and PTBP1 knockout samples were subjected to SDS-PAGE. ab133734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)

      Immunohistochemical analysis of PTBP1 in paraffin embedded Human breast carcinoma tissue stained with ab133734 at a 1/100 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Western blot - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      All lanes : Anti-PTBP1 antibody [EPR9048(B)] (ab133734) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : PTBP1 knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 57 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab133734 observed at 57 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab133734 was shown to recognize PTBP1 in wild-type HAP1 cells as signal was lost at the expected MW in PTBP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PTBP1 knockout samples were subjected to SDS-PAGE. Ab133734 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Western blot - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      All lanes : Anti-PTBP1 antibody [EPR9048(B)] (ab133734) at 1/10000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : Daudi cell lysate
      Lane 4 : A549 cell lysate
      Lane 5 : BxPC3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

      Predicted band size: 57 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Immunocytochemistry/ Immunofluorescence - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Immunofluorescent staining of PTBP1 in A549 cells, using ab133734 at a 1/250 dilution.
    • Flow Cytometry - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)
      Flow Cytometry - Anti-PTBP1 antibody [EPR9048(B)] (ab133734)

      Flow cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling PTBP1 (red) with purified ab133734 at a 1/2000 dilution (1ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

    References

    Publishing research using ab133734? Please let us know so that we can cite the reference in this datasheet.

    ab133734 has been referenced in 6 publications.

    • Urdaneta EC  et al. Purification of cross-linked RNA-protein complexes by phenol-toluol extraction. Nat Commun 10:990 (2019). PubMed: 30824702
    • Jiang D  et al. Long noncoding RNA HCG22 suppresses proliferation and metastasis of bladder cancer cells by regulation of PTBP1. J Cell Physiol N/A:N/A (2019). PubMed: 31304601
    • Sayed ME  et al. NOVA1 directs PTBP1 to hTERT pre-mRNA and promotes telomerase activity in cancer cells. Oncogene 38:2937-2952 (2019). PubMed: 30568224
    • Dumbovic G  et al. A novel long non-coding RNA from NBL2 pericentromeric macrosatellite forms a perinucleolar aggregate structure in colon cancer. Nucleic Acids Res 46:5504-5524 (2018). PubMed: 29912433
    • Yuan T  et al. Low-density lipoprotein receptor-related protein 6 regulates alternative pre-mRNA splicing. J Cell Mol Med 22:4653-4663 (2018). PubMed: 30070011
    • Hou W  et al. Two Polypyrimidine Tracts in Intron 4 of the Major Immediate Early Gene Are Critical for Gene Expression Switching from IE1 to IE2 and for Replication of Human Cytomegalovirus. J Virol 90:7339-49 (2016). ChIP ; Human . PubMed: 27252533

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