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Anti-Perforin 抗体 [B-D48] - BSA and Azide free (ab47225)

Reviews (1)Q&A (4)References (18)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Perforin antibody [B-D48] - BSA and Azide free (ab47225)

    Key features and details

    • Mouse monoclonal [B-D48] to Perforin - BSA and Azide free
    • Suitable for: IHC-P
    • Reacts with: Human
    • Isotype: IgG1

    Conjugates logo Related conjugates and formulations

    FITC PE

    製品の概要

    • 製品名

      Anti-Perforin antibody [B-D48] - BSA and Azide free
      Perforin 一次抗体 製品一覧

    • 製品の詳細

      Mouse monoclonal [B-D48] to Perforin - BSA and Azide free

    • 由来種

      Mouse

    • アプリケーション

      適用あり: IHC-Pmore details

    • 種交差性

      交差種: Human

    • 免疫原

      Recombinant human perforin.

    • ポジティブ･コントロール

      • IHC-P: Human spleen FFPE tissue sections.

    • 特記事項

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    製品の特性

    • 製品の状態

      Liquid

    • 保存方法

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

    • バッファー

      pH: 7.30
      Constituent: 100% PBS

      Sterile-filtered through 0.22 µm. Carrier and preservative free

    • キャリア・フリー

      はい
    • Concentration information loading...

    • 精製度

      Ion Exchange Chromatography

    • 特記事項（精製）

      Recognizes human Perforin, a 70 kDa protein.

    • ポリ/モノ

      モノクローナル

    • クローン名

      B-D48

    • ミエローマ

      x63-Ag8.653

    • アイソタイプ

      IgG1

    • 軽鎖の種類

      kappa

    • 研究分野

    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ab47225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
    IHC-P (1)
    Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
    特記事項
    IHC-P
    Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    ターゲット情報

    • 機能

      Plays a key role in secretory granule-dependent cell death, and in defense against virus-infected or neoplastic cells. Plays an important role in killing other cells that are recognized as non-self by the immune system, e.g. in transplant rejection or some forms of autoimmune disease. Can insert into the membrane of target cells in its calcium-bound form, oligomerize and form large pores. Promotes cytolysis and apoptosis of target cells by facilitating the uptake of cytotoxic granzymes.

    • 関連疾患

      Defects in PRF1 are the cause of hemophagocytic lymphohistiocytosis familial type 2 (FHL2) [MIM:603553]; also known as HPLH2. Familial hemophagocytic lymphohistiocytosis (FHL) is a genetically heterogeneous, rare autosomal recessive disorder. It is characterized by immune dysregulation with hypercytokinemia and defective natural killer cell function. The clinical features of the disease include fever, hepatosplenomegaly, cytopenia, hypertriglyceridemia, hypofibrinogenemia, and neurological abnormalities ranging from irritability and hypotonia to seizures, cranial nerve deficits, and ataxia. Hemophagocytosis is a prominent feature of the disease, and a non-malignant infiltration of macrophages and activated T lymphocytes in lymph nodes, spleen, and other organs is also found.

    • 配列類似性

      Belongs to the complement C6/C7/C8/C9 family.
      Contains 1 C2 domain.
      Contains 1 EGF-like domain.
      Contains 1 MACPF domain.

    • ドメイン

      The C2 domain mediates calcium-dependent binding to lipid membranes. A subsequent conformation change leads to membrane insertion of beta-hairpin structures and pore formation. The pore is formed by transmembrane beta-strands.

    • 翻訳後修飾

      N-glycosylated.

    • 細胞内局在

      Cytoplasmic granule lumen. Secreted. Cell membrane. Endosome lumen. Stored in cytoplasmic granules of cytolytic T-lymphocytes and secreted into the cleft between T-lymphocyte and target cell. Inserts into the cell membrane of target cells and forms pores. Membrane insertion and pore formation requires a major conformation change. May be taken up via endocytosis involving clathrin-coated vesicles and accumulate in a first time in large early endosomes.
    • Information by UniProt

    • 参照データベース

      • 別名

        • Cytolysin antibody
        • FLH2 antibody
        • HPLH2 antibody
        • Lymphocyte pore-forming protein antibody
        • P1 antibody
        • PERF_HUMAN antibody
        • perforin 1 (pore forming protein) antibody
        • Perforin 1 antibody
        • Perforin-1 antibody
        • PFP antibody
        • PGFL antibody
        • PIGF antibody
        • PIGF-2 antibody
        • PLGF antibody
        • Pore forming protein antibody
        • prf1 antibody
        • SHGC-10760 antibody
        see all

      画像

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Perforin antibody [B-D48] - BSA and Azide free (ab47225)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Perforin antibody [B-D48] - BSA and Azide free (ab47225)

        IHC image of Perforin staining in human spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. 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 ab47225, 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.

      参考文献 (18)

      ab47225 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。

      ab47225 は 18 報の論文で使用されています。

      • Vitanza NA  et al. Intraventricular B7-H3 CAR T Cells for Diffuse Intrinsic Pontine Glioma: Preliminary First-in-Human Bioactivity and Safety. Cancer Discov 13:114-131 (2023). PubMed: 36259971
      • Mijnheer G  et al. Compartmentalization and persistence of dominant (regulatory) T cell clones indicates antigen skewing in juvenile idiopathic arthritis. Elife 12:N/A (2023). PubMed: 36688525
      • Velasco Cárdenas RM  et al. Harnessing CD3 diversity to optimize CAR T cells. Nat Immunol 24:2135-2149 (2023). PubMed: 37932456
      • Schwabenland M  et al. Deep spatial profiling of human COVID-19 brains reveals neuroinflammation with distinct microanatomical microglia-T-cell interactions. Immunity 54:1594-1610.e11 (2021). PubMed: 34174183
      • Arunachalam PS  et al. Systems vaccinology of the BNT162b2 mRNA vaccine in humans. Nature 596:410-416 (2021). PubMed: 34252919
      View all Publications for this product

      レビューと Q&A

      Show All レビュー Q&A
      レビューを送る 質問を送る

      1-5 of 5 Abreviews or Q&A

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Perforin antibody [B-D48] - BSA and Azide free

       Good
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Human Tissue sections (Tonsil)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: 10mM Tris Base, 1mM EDTA, pH 9.2
      Permeabilization
      No
      Specification
      Tonsil
      Blocking step
      3% BSA, 5% horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 8% · 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 May 30 2022

      Question


      Thank you so much for such a detailed response. This will help us moving
      forward. Do you know if this particular clone cross-reacts with non-human
      primates?

      Read More
      Answer

      You are welcome.

      We do not have any data for cross-reactivity with non-primate samples. I tried finding an amino acid sequence for non-primate perforin to align with human perforin, which was the immunogen for ab47225.

      The unreviewed UniProt entry for a species of macaque,

      http://www.uniprot.org/uniprot/B3FWV1

      is 95% identical with the sequence of human perforin

      (http://www.uniprot.org/uniprot/P14222),

      which suggests that, at least for this species, cross-reactivity is likely.

      Read More

      Question

      No staining by IHC. We are staining Human rectal biopsy pieces that have been fixed in 4%PFA followed by 70% Ethanol before embedding in parrafin and cutting sections
      at 10 microns. We then use a 0.01M Citrate Buffer pH 6.0 for antigen retrieval. We use the recommended 20ug/ml in 5%BSA/TBS for one hour at room temperature.

      Read More
      Answer

      Thank you for sending the details of your protocol.

      I think for PFA-fixed, paraffin-embedded tissue, your protocol is correct. However, our source for this clone has told us that it was only tested in the past for IHC on frozen tissue by collaborators, and they did not provide a protocol. I suspect they may have used a coagulative fixative such as acetone or methanol. These are typically used to preserve tissue before freezing, as they do not introduce chemical modification to proteins, unlike formaldehyde-based fixatives. These modification tend to interfere with antibody:antigen recognition. The retrieval step aims to reverse this, but is not always effective.

      I can suggest trying an enzymatic retrieval instead of a heat-mediated retrieval (trypsin or proteinase K), but we have no data indicating this will or will not work. I do suggest obtaining a positive control of spleen tissue, to confirm that the results are negative due to the procedure or antibody, and not due to lack of expression.

      Read More

      Question


      I am using the Abcam Anti-perforin antibody [B-D48] ab47225 for IHC
      staining on human rectal tissue. I am having some difficulty in detection
      of perforin and was wondering if you could tell me what antigen retrieval
      and permeabilization method was used in the testing of this antibody for
      IHC-Fr usage.

      Read More
      Answer

      Thank you for bringing this to our attention. To our knowledge, no antigen retrieval should be necessary for histochemistry on frozen tissue sections, or permeabilization assuming your sections are no thicker than 10 microns.

      Can you please tell me what your tissue samples are (tissue type and species), how it is fixed, and how much antibody you are incubating with the sections and for how long? Are you confident the secondary antibody is effective with mouse IgG? If you have an image of the staining, could you please send it?

      I look forward to your reply.

      Read More

      Question

      I am interested in ordering: anti-human Perforin (Cat. No. ab47225) purified Before I order this antibody, however, I need to ensure that it is free of any and all carrier proteins, i.e. BSA or gelatin. Can you please confirm this for me?

      Read More
      Answer

      Thank you for contacting us. I have doublechecked with the lab and they have confirmed that the antibody ab47225 is provided just in PBS, without any carrier protein or preservative. Format: Phosphate-buffered saline. Sterile-filtered through 0.22 µm. Carrier and preservative free. 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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