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リコンビナントRabMAb

Anti-PAP 抗体 [EPR4067] (ab109004)

Reviews (1)Q&A (8)References (1)
Western blot - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunocytochemistry/ Immunofluorescence - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
  • Western blot - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
  • Anti-PAP antibody [EPR4067] (ab109004)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4067] to PAP
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-PAP antibody [EPR4067]
    PAP 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR4067] to PAP

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IHC-P, ICC/IFmore details
    適用なし: Flow Cyt or IP

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

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

  • 特記事項

    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.

    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.

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

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab109004の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
1/1000 - 1/10000. Predicted molecular weight: 45 kDa.
IHC-P (1)
1/40000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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/IF
1/50 - 1/100.
特記事項
WB
1/1000 - 1/10000. Predicted molecular weight: 45 kDa.
IHC-P
1/40000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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/IF
1/50 - 1/100.
追加情報
Is unsuitable for Flow Cyt or IP.

ターゲット情報

  • 配列類似性

    Belongs to the histidine acid phosphatase family.

  • 細胞内局在

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

  • 参照データベース

    • 別名

      • 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

    画像

    • Western blot - Anti-PAP antibody [EPR4067] (ab109004)
      Western blot - Anti-PAP antibody [EPR4067] (ab109004)
      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 - Anti-PAP antibody [EPR4067] (ab109004)
      Immunocytochemistry/ Immunofluorescence - Anti-PAP antibody [EPR4067] (ab109004)

      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) - Anti-PAP antibody [EPR4067] (ab109004)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)

      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.

    • Western blot - Anti-PAP antibody [EPR4067] (ab109004)
      Western blot - Anti-PAP antibody [EPR4067] (ab109004)
      Anti-PAP antibody [EPR4067] (ab109004) at 1/1000 dilution ((unpurified)) + Prostate cancer lysate at 10 µg

      Predicted band size: 45 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAP antibody [EPR4067] (ab109004)

      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.

    • Anti-PAP antibody [EPR4067] (ab109004)
      Anti-PAP antibody [EPR4067] (ab109004)

    参考文献 (1)

    ab109004 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab109004 は 1 報の論文で使用されています。

    • Kirschenbaum A  et al. Prostatic Acid Phosphatase Alters the RANKL/OPG System and Induces Osteoblastic Prostate Cancer Bone Metastases. Endocrinology 157:4526-4533 (2016). PubMed: 27783536

    レビューと Q&A

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    1-9 of 9 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Prostatic Acid Phosphatase antibody [EPR4067]

     Good
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Dec 31 2018

    Question


    Thank you for your answer,

    Regarding your questions:

    1) yes, our secondary antibody is working with other rabbit primary antibodies.

    2) I will try an BSA for blocking and I will let you know if this works.

    Best regards

    Read More
    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

    Inquiry: Replacement or refund for product 1) Abcam product code ab109004 2) Abcam order reference number or product batch number GR41390-5 3) Description of the problem: The antibody does not detect human nor mouse PAP. 4) Sample preparation: Type of sample: whole cell lysates C4-2 prostate cancer cells mouse skin, mouse prostate Lysis buffer: RIPA Protease inhibitors: cocktail Phosphatase inhibitors: yes Reducing agent: yes Boiling for ≥5 min? yes Protein loaded 30-50ug/lane Positive control: any suggestion? Negative control: any suggestion? 5) Percentage of gel: 4-12% Type of membrane: PVDF Protein transfer verified: yes Blocking agent and concentration: 5%milk PBS-T Blocking time: 2h Blocking temperature: RT 6) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution: 1/200 and 1/500 Diluent buffer: PBS-T 1% milk Incubation time: 12h Incubation temperature: 4° 7) Secondary antibody: GAR HRP Concentration or dilution: 1:10000 Diluent buffer: PBS-T 1% milk Incubation time: 1h Incubation temperature: RT Fluorochrome or enzyme conjugate: HRP 8) Washing after primary and secondary antibodies: Buffer: PBS-T Number of washes: 3 9)Detection method: luminesence SuperSignal West Dura 10) How many times have you run this staining? 3 Do you obtain the same results every time? yes What steps have you altered to try and optimize the use of this antibody? concentration of primary antibody, type of sample (mouse tissu)

    Read More
    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/HRP-GAPDH-antibody-Loading-Control-ab9385.html (or use the following: https://www.abcam.com/HRP-GAPDH-antibody-Loading-Control-ab9385.html).

    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

    Hi, Order number is Order #*** (PO:REF) Thanks Eriks

    Read More
    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

    Question

    Hi, I gave second chance for this antibody, this tame I tried mouse testis and kidney - still nothing. I give up. I looking for good anti-phosphotyrosine antibody in mouse. See Abcam have one Anti-Phosphotyrosine antibody (ab9319). So can I change ACPP antibody to ab9319? Thanks

    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

    I mean - others antiACPP antibodies? What about others antibodies what you have in Abcam?

    Read More
    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

    Question

    Hi, Recently we bought from Abcam Anti-Prostatic Acid Phosphatase antibody [EPR4067] (ab109004). I tried to westernblot mouse liver total lysate to see ACPP (Prostatic Acid Phosphatase) expression in liver (as RT-qPCR show fairy well expression there – Ct value is ~25) and can not see it at all. As documentation say – it react with mouse ACPP. Did company tried to blot any real mouse tissues? Thank you,    

    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

    Question

    I just ordered this antibody. But still have one question - against which synthetic peptide sequence antibody is made. As we know there is 2 types of ACPP in mammalian cells. So antibody recognize both?

    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

    Question

    Hi, I would like to test Anti-Prostatic Acid Phosphatase antibody [EPR4067] (ab109004) on my mutant mice as mice ACPP mRNA is low if compare with wilt type. As Abcam websate say –  ab109004 antibody reacts with mouse ACPP, is it correct? Thank you!

    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

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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