Overview

  • Product name
  • Description
    Goat polyclonal to PTBP1
  • Host species
    Goat
  • Tested applications
    Suitable for: IP, IHC-P, ICC/IF, WB, ELISAmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Cow, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to Human PTBP1 aa 2-14 (N terminal).
    Sequence:

    DGIVPDIAVGTKR-C


    Database link: P26599
    (Peptide available as ab23105)

  • Positive control
    • Jurkat lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab5642 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. PubMed: 17967866
IHC-P Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 0.1 - 0.3 µg/ml. Detects a band of approximately 60-65 kDa (predicted molecular weight: 61-64 kDa).Can be blocked with Human PTBP1 peptide (ab23105).
ELISA Use at an assay dependent concentration.

Antibody detection limit dilution 1:32,000.

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

  • All lanes : Anti-PTBP1 antibody (ab5642) at 0.01 µg/ml

    Lane 1 : Human epithelial cells from cervix adenocarcinoma
    Lane 2 : Human liver hepatocellular carcinoma
    Lane 3 : Human epithelial cells from embryonic kidney

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 61-64 kDa

  • Immunohistochemical analysis of paraffin embedded Human Kidney tissue labeling PTBP1 with ab5642 at 2 ug/ml. Tissue underwent antigen retrieval in steam with Tris/EDTA buffer (pH 9.0). The HRP-staining procedure was used for detection.

  • Immunohistochemical analysis of paraffin embedded Human Kidney tissue labeling PTBP1 with ab5642 at 2 ug/ml. Tissue underwent antigen retrieval in steam with citrate buffer (pH 6.0). The HRP-staining procedure was used for detection.

  • ab5642 staining (1µg/ml) of Jurkat lysate (RIPA buffer, 30µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence.

    ab5642 staining (1µg/ml) of Jurkat lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
  • ICC/IF image of ab5642 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5642, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab5642 staining in human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5642, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Anti-PTBP1 antibody (ab5642) at 0.1 µg/ml + Human Ovary Lysate in RIPA buffer at 35 µg

    Developed using the ECL technique.

    Predicted band size: 61-64 kDa

  • ab5642 (5µg/ml) staining of paraffin embedded Human Skin. Steamed antigen retrieval with citrate buffer pH 6, AP-staining shows nuclear staining of keratinocytes.

References

This product has been referenced in:
  • Georgilis A  et al. PTBP1-Mediated Alternative Splicing Regulates the Inflammatory Secretome and the Pro-tumorigenic Effects of Senescent Cells. Cancer Cell 34:85-102.e9 (2018). Read more (PubMed: 29990503) »
  • Aguilera O  et al. Vitamin C uncouples the Warburg metabolic switch in KRAS mutant colon cancer. Oncotarget 7:47954-47965 (2016). Read more (PubMed: 27323830) »
See all 9 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for the details that you have provided. At this point, I would like to make the following suggestions. Use ab5642 at a concentration of 1 µg/ml and incubate overnight at 4C. Do not re-use the primary antibody. Ensure that your secondary antibody is working properly, and you may need to increase the concentration of the secondary antibody as well as the incubation period. Do not overwash the membrane. Also, I suggest running a positive control - Jurkat lysate is recommended as positive control for ab5642.

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Answer

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab5642. A secondary antibody that is compatible for use with this primary is ab6741 - Rabbit polyclonal to Goat IgG H&L (HRP). I would like to help you out and if you would kindly answer the questions below, the answers will enable us to investigate this matter as quickly as possible. Also, please include the batch number that you received (it is located on the vial) and the Abcam order number or purchase order number that was used. 1. Please describe the problem (high background, wrong band size, more bands, no band etc). 2. On what material are you testing the antibody in WB? •Species? •Cell extract/ Nuclear extract? •Purified protein? •Recombinant protein? 3. How much protein did you load? •How did you prepare the lysate for the analysis (protease inhibitors etc)? •Did you heat the samples? 4. Primary Antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash step? 5. Secondary Antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash step? •Do you know whether the problems you are experiencing come from the secondary? 6. What detection method are you using? 7. Background bands •Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control) •Is the blocking step sufficient? (We recommend blocking the membrane by adding 20 ml of blocking buffer (5% non-fat dry milk, 0.1% Tween-20 in TBS). Incubate for 2 h at room temperature or overnight at 4°C with agitation) •Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) •At what size are the bands migrating? Could they be degradation products of your target? •Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 8. Optimization attempts •How many times have you tried the Western? •Do you obtain the same results every time e.g. are background bands always in the same place? •What steps have you altered? 9. Did you apply positive and negative controls along with the samples? Please specify.

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