Recombinant
RabMAb

Recombinant Anti-PAP antibody [EPR4067] (ab109004)

Overview

  • Product name

    Anti-PAP antibody [EPR4067]
    See all PAP primary antibodies
  • Description

    Rabbit monoclonal [EPR4067] to PAP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICCmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PAP aa 300-400. The exact sequence is proprietary.

  • Positive control

    • WB: Human prostate lysates. IHC-P: human prostate and adenocarcinoma tissue. ICC/IF: PC-3 cells.
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

     

     This product was previously labelled as Prostatic Acid Phosphatase

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109004 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 1/1000 - 1/10000. Predicted molecular weight: 45 kDa.
IHC-P 1/40000.

Heat up to 98 &degC, below boiling, and then let cool for 10-20 min.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500

ICC 1/50 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Sequence similarities

      Belongs to the histidine acid phosphatase family.
    • Cellular localization

      Secreted and Lysosome membrane. Predominantly localized in the plasma membrane but also detected in intracellular vesicles.
    • Information by UniProt
    • Database links

    • Alternative names

      • 5' NT antibody
      • 5' nucleotidase antibody
      • 5'-NT antibody
      • 5'-nucleotidase antibody
      • Acid phosphatase prostate antibody
      • ACP 3 antibody
      • ACP3 antibody
      • acpP antibody
      • Ecto 5' nucleotidase antibody
      • Ecto-5'-nucleotidase antibody
      • PAP antibody
      • PPAP_HUMAN antibody
      • Prostatic acid phosphatase antibody
      • Prostatic acid phosphotase antibody
      • Thiamine monophosphatase antibody
      • TMPase antibody
      see all

    Images

    • Anti-PAP antibody [EPR4067] (ab109004) at 1/1000 dilution (Purified) + Human prostate lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 45 kDa
      Observed band size: 50 kDa
      why is the actual band size different from the predicted?

    • Immunocytochemistry/ Immunofluorescence analysis of PC-3 (Human prostate adenocarcinoma epithelial cell) cells labeling PAP with purified ab109004 at 1/50 dilution (1.92 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate tissue sections labeling PAP with purified ab109004 at 1/40,000 dilution (0.0024 µg/ml). Heat mediated antigen retrieval using Bond� Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Anti-PAP antibody [EPR4067] (ab109004) at 1/1000 dilution ((unpurified)) + Prostate cancer lysate at 10 µg

      Predicted band size: 45 kDa

    • Unpurified ab109004 at 1/250 dilution, staining PAP in Human prostatic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.

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

    • Unpurified ab109004 at 1/250 dilution, staining PAP in Human prostatic tissue by Immunohistochemistry, Paraffin-embedded tissue.

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

    • Unpurified ab109004 showing negative staining in Breast carcinoma tissue.

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

    • Unpurified ab109004 showing negative staining in Normal uterus tissue.

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

    • Unpurified ab109004 showing positive staining in Prostate hyperplasia tissue.

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

    References

    ab109004 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-9 of 9 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Prostate)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    No
    Specification
    Prostate
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 31 2018

    Answer

    Thank you very much for your reply.

    I hope this will resolve the problem - please keep us updated!

    I wish you a good day and look forward hearing back from you!

    Read More

    Question
    Answer

    Thank you for contacting us. I am very sorry to hear that you have been experiencing problems also with this antibody.

    I have looked through the protocol provided and compared it to the information we have. There are only two points which I would ask you to double check: 1.) Do you know whether the secondary antibody is working with other rabbit primary antibodies? 2.) I can also recommend to test another alternative blocking buffer, such as BSA for blocking. Indeed, changing the blocking buffer can significantly improve the results. As an illustration for this, please see the Western blot image on the datasheet of ab9385 https://www.abcam.com/index.html?datasheet=9385 (or use the following: https://www.abcam.com/index.html?datasheet=9385).

    If the antibody is faulty,I would be pleased to arrange also for a refund or a replacement antibody for you for this antibody.

    Thank you for your cooperation. I am looking forward to hear back from you.

    Read More

    Question
    Answer

    Thank you for your reply. I am sorry that the results with ab109004 were so poor. I am sending a free of charge vial of ab9319 on the order# ***, which should arrive tomorrow. Please keep me updated about the results with this replacement antibody, and let me know if you have any questions or if there is anything else that we can do for you.

    Read More

    Answer

    Sure, I can send a vial of ab9319. Could you send me your order or PO number so I can find your original order?

    Read More

    Question
    Answer

    Thank you for your reply. I would be happy to send an antibody to replace the vial of ab109004, but unfortunately we don't have any other ACPP antibodies that have been validated in mouse. So we can not guarantee that another ACPP antibody will work with mouse samples. Is there a different antibody that you would like to try? I look forward to hearing from you. Please let me know if you have any further questions.

    Read More

    Answer

    Thank you for contacting us and letting us know about the trouble with this antibody. Ab109004 has tested positive in Western blot with murine lysate, but I do have a few additional questions so that I can help further. 1) Are there any non-specific bands on the blot, or are there no bands at all? Are you able to provide an image? 2) What dilution of antibody do you use, and how long do you incubate the membrane with the antibody? 3) What blocking solution is used (BSA, milk, etc)? 4) How much protein was loaded per lane of the gel? 5) Some sources indicate that ACPP is not expressed in healthy liver (see Human Protein Atlas link, below) though this conflicts with your qRT-PCR data. Do you have a positive control lysate, such as prostate or prostate cancer lyste, that you could run in parallel with the liver samples? http://www.proteinatlas.org/ENSG00000014257 I look forward to hearing from you. We do guarantee all of our products to work as stated on the datasheet for up to 6 months after purchase, so if we can't get the antibody to work I will be happy to send a replacement or issue a credit or refund. Please let me know if you have any questions or if there is anything else that I can do for you.

    Read More

    Answer

    The immunogen peptide maps between amino acids 325 and 355 of human Prostatic Acid Phosphatase (UniProt: P15309). The antibody is predicted to recognize both isoforms of ACPP, based on the immunogen. Please contact us if you have any other questions.

    Read More

    Answer

    Thank you for contacting us. This product has been tested successfully in human and mouse in WB, IP, IHC-P, ICC applications. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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