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Anti-Perforin抗体[CB5.4] (ab16074)

Reviews (5)Q&A (5)References (31)

Key features and details

  • Rat monoclonal [CB5.4] to Perforin
  • Suitable for: IP, ICC, WB, IHC-Fr, IHC-P, ICC/IF
  • Reacts with: Mouse
  • Isotype: IgG2a

概述

  • 产品名称

    Anti-Perforin抗体[CB5.4]
    参阅全部 Perforin 一抗

  • 描述

    大鼠单克隆抗体[CB5.4] to Perforin

  • 宿主

    Rat

  • 经测试应用

    适用于: IP, ICC, WB, IHC-Fr, IHC-P, ICC/IFmore details

  • 种属反应性

    与反应: Mouse

  • 免疫原

    Recombinant fragment, corresponding to amino acids 98 - 534 of Mouse Perforin.

  • 表位

    This antibody recognizes mouse perforin, region amino acids 402-534.

  • 常规说明

    This product was changed from ascites to tissue culture supernatant on 31st May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab16074于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
IP
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
WB (1)
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IHC-P (3)
Use at an assay dependent concentration.
ICC/IF (1)
Use at an assay dependent concentration.
说明
IP
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

靶标

  • 功能

    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.

  • 疾病相关

    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.

  • 序列相似性

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

  • 结构域

    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.

  • 翻译后修饰

    N-glycosylated.

  • 细胞定位

    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

  • 数据库链接

    • 别名

      • 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

    文献 (31)

    发表研究结果有使用 ab16074？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab16074 被引用在 31 文献中.

    • Wang Z  et al. Sphingomyelin-derived nanovesicles for the delivery of the IDO1 inhibitor epacadostat enhance metastatic and post-surgical melanoma immunotherapy. Nat Commun 14:7235 (2023). PubMed: 37945606
    • Qiu C & Du G Loss of LEF-1 expression as a diagnostic indicator for extranodal NK/T-cell lymphoma: An immunohistochemical study of 88 cases. Eur J Haematol 109:513-518 (2022). PubMed: 35871391
    • Ravindran Menon D  et al. EGCG Inhibits Tumor Growth in Melanoma by Targeting JAK-STAT Signaling and Its Downstream PD-L1/PD-L2-PD1 Axis in Tumors and Enhancing Cytotoxic T-Cell Responses. Pharmaceuticals (Basel) 14:N/A (2021). PubMed: 34832863
    • Liu X  et al. Development of Facile and Versatile Platinum Drug Delivering Silicasome Nanocarriers for Efficient Pancreatic Cancer Chemo-Immunotherapy. Small 17:e2005993 (2021). PubMed: 33682329
    • Liu X  et al. Combination Chemo-Immunotherapy for Pancreatic Cancer Using the Immunogenic Effects of an Irinotecan Silicasome Nanocarrier Plus Anti-PD-1. Adv Sci (Weinh) 8:2002147 (2021). PubMed: 33747719
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-Perforin antibody [CB5.4]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (CT26 cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin gel)
    Loading amount
    40 µg
    Specification
    CT26 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    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

    提交于 Mar 08 2022

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Perforin antibody [CB5.4]

     Poor
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (Small intestinal LGL lymphoma (confirmed case))
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer pH6.0
    Permeabilization
    No
    Specification
    Small intestinal LGL lymphoma (confirmed case)
    Blocking step
    Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Formaldehyde
    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.

    Ikki Mitsui

    Verified customer

    提交于 Aug 28 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Perforin antibody [CB5.4]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    spleen
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
    Fixative
    10% NBF
    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

    提交于 Jul 27 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Perforin antibody [CB5.4]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Sample
    Mouse Cell (uterus, placenta)
    Specification
    uterus, placenta
    Permeabilization
    No
    Fixative
    10% neutral buffered formalin
    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.

    DR. Varkha Agrawal

    Verified customer

    提交于 Jul 03 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Perforin antibody [CB5.4]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Lymphoid tissue)
    Specification
    Lymphoid tissue
    Fixative
    Zinc
    Antigen retrieval step
    None
    Permeabilization
    No
    Blocking step
    Rodent block-M (other company) as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
    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

    提交于 Jan 28 2009

    Question

    Phone call requesting information on which antigen retrieval would be recommended to use the Anti-Perforin antibody [CB5.4] (ab16074).

    Read More
    Answer

    I can confirm that the Anti-Perforin antibody [CB5.4] (ab16074) has been used with citrate buffer (pH 6) when performing staining of paraffin embedded sections. I would however advise that it may be worthwhile to optimize this for your particular tissue preparations. Therefore using both citrate buffer (pH 6) as well as Tris-EDTA (pH 9) and seeing which provides the best results for you.

    Read More

    Question

    ein kostenloser Antikörper wäre mir lieber. Wenn es möglich ist, dass ich mir diesen aussuchen kann, dann würde ich mich für folgenden entscheiden: https://www.abcam.com/F4-80-antibody-CI-A3-1-Allophycocyanin-ab105080.html.

    Read More
    Answer

    Vielen Dank für Ihre Anfrage.

    Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 123456 und solltemorgen bei Ihnen ankommen.

    Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Question

    Hier das ausgefüllte questionnaire, im Anhang die Bilder (nur der rote Kanal =Perforin):

    2nd rab rat ist das Bild der Kontrollfärbung, wo nur 2. Antikörper gefärbt wurden. Die Zahlen Perf-50 heißt, dass der Primärantikörper gegen Perforin in einer Verdünnung von 1:50 verwendet wurde.

    Das Bild 12-08-07 spleen perf 50 cd8 50 stammt aus der ersten Färbung (Titration 1) die beiden anderen aus der 2. Färbung



    1) Abcam product code ab 16074



    2) Abcam order reference number or product batch number



    3) Description of the problem

    not sure whether the signal is just background, since control without primary antibodies but with secondary antibodies also gives a signal (see attached pictures)



    4) Sample preparation:

    Species mouse, spleen embedded in tissue tek, frozen at -80°C

    Type of sample: Fresh frozen sections (5µm)

    Sample preparation: fixed in acetone for 10 min at RT, encirceled with DAKO pen, blocked with serum, incubation with primary antibodies, washing, incubation with secondary antibodies, washing, incubation with Hoechst, washing, cover slipping with Glycerin/Gelatine

    Positive control staining of spleen

    Negative control only secondary antibodies



    5) Fixation step

    Yes

    If yes: fixed in acetone

    Fixation time10 min

    Fixation temperature RT



    6) Antigen retrieval method

    none



    7) Permeabilization method: no

    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

    Permeabilizing agent and concentration:





    8) Blocking agent (eg BSA, serum…): serum (goat or chicken, depending on the second antibody)

    Concentration 1/100

    Blocking time at least 10 min

    Blocking temperature RT



    9) Endogenous peroxidases blocked? no

    Endogenous biotins blocked? no



    10) Primary antibody (If more than one was used, describe in “additional notes”) :

    Concentration or dilution 1:10, 1:20, 1:50, 1:50, 1:500, 1:1000

    Diluent buffer PBS

    Incubation time 1 hour



    11) Secondary antibody: anti-rat Alexa 594 (invitrogen A21471)

    Species:

    Reacts against: rat

    Concentration or dilution 1:200

    Diluent buffer PBS

    Incubation time 1 hour

    Fluorochrome or enzyme conjugate Alexa 594



    12) Washing after primary and secondary antibodies:

    Buffer: PBS

    Number of washes 2 times in a PBS bath



    13) Detection method



    14) How many times have you run this staining? 2 titrations 1.: 1:50, 1:500, 1:1000, 2.: 1:10, 1:20, 1;50

    Do you obtain the same results every time? no

    What steps have you altered to try and optimize the use of this antibody?

    altered washing steps for 2nd titration (did not dip slides into PBS bath but dribbled PBS on top and tapped it off again, also 2 times



    Falls Sie noch etwas benötigen, werde ich es Ihnen zukommen lassen.



    Vielen Dank für Ihre Hilfe,

    Read More
    Answer

    Vielen Dank für Ihre Antwort und für diese weiteren Informationen.

    Da Ihrem Protokoll bis auf Kleinigkeiten nichts hinzuzufügen ist, muss ich leider davon ausgehen, dass der Antikörper nicht so funktioniert hat, wie er sollte.

    Ich weiß, dass Sie viel Zeit für diese Experimente verwendet haben und möchte Ihnen daher einen kostenlosen Antikörperoder einen Gutschein als Ersatz anbieten. Würden Sie mir bitte mitteilen, welche Alternative Sie bevorzugen?

    Vielen Dank für Ihre Kooperation und Hilfsbereitschaft. Ich freue mich bald wieder von Ihnen zu hören.

    Read More

    Question

    Concentration is way too high in IHC-Fr (1/10).
    Isotyp controll?

    Read More
    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid zu hören, dass Sie Probleme mit diesem Antikörper haben.

    Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.



    Ich freue mich, bald wieder von Ihnen zu hören.

    Read More

    Question

    ICC/IF with mouse T-cells and B-cells T-cells show some staining, but B-cells as well (they should not show staining) IgG control: showed no stainig, thus secondary Ab is fine Ab: 5 ug/ml block w: none fix/perm: with commercialized Cytofix/perm solution

    Read More
    Answer

    Thank you for contacting us. Your credit note ID is xxx. I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

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