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    perforin-antibody-cb54-ab16074.pdf

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Immunology Adaptive Immunity T Cells Cytotoxic Cells
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Anti-Perforin antibody [CB5.4] (ab16074)

  • Datasheet
  • SDS
Reviews (4)Q&A (5)References (22)

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Key features and details

  • Rat monoclonal [CB5.4] to Perforin
  • Reacts with: Mouse
  • Isotype: IgG2a

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Protein
Product image
Recombinant Human Perforin protein (ab114201)
Secondary
Product image
Goat Anti-Rat IgG H&L (Alexa Fluor® 488) (ab150157)
Primary
Product image
Anti-PD1 antibody [EPR20665] (ab214421)

View more associated products

Overview

  • Product name

    Anti-Perforin antibody [CB5.4]
    See all Perforin primary antibodies
  • Description

    Rat monoclonal [CB5.4] to Perforin
  • Host species

    Rat
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

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

  • Epitope

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

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    CB5.4
  • Isotype

    IgG2a
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Cytotoxic Cells
    • Immunology
    • Innate Immunity
    • Complement
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Rat IgG H&L (Alexa Fluor® 488) (ab150157)
    • Goat Anti-Rat IgG H&L (HRP) (ab205720)
  • Isotype control

    • Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (ab18450)
  • Recombinant Protein

    • Recombinant Human Perforin protein (ab114201)

Target

  • Function

    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.
  • Involvement in disease

    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.
  • Sequence similarities

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

    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.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    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.
  • Target information above from: UniProt accession P14222 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 18646 Mouse
    • SwissProt: P10820 Mouse
    • Unigene: 240313 Mouse
    • Alternative names

      • 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

    Protocols

    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (22)

    Publishing research using ab16074? Please let us know so that we can cite the reference in this datasheet.

    ab16074 has been referenced in 22 publications.

    • Choi YH  et al. IL-27 enhances IL-15/IL-18-mediated activation of human natural killer cells. J Immunother Cancer 7:168 (2019). PubMed: 31277710
    • Wijesundara DK  et al. Induction of Genotype Cross-Reactive, Hepatitis C Virus-Specific, Cell-Mediated Immunity in DNA-Vaccinated Mice. J Virol 92:N/A (2018). PubMed: 29437963
    • Lu J  et al. Breast Cancer Chemo-immunotherapy through Liposomal Delivery of an Immunogenic Cell Death Stimulus Plus Interference in the IDO-1 Pathway. ACS Nano 12:11041-11061 (2018). PubMed: 30481959
    • Lu J  et al. Nano-enabled pancreas cancer immunotherapy using immunogenic cell death and reversing immunosuppression. Nat Commun 8:1811 (2017). PubMed: 29180759
    • Kenchegowda D  et al. Inactivation of maternal Hif-1a at mid-pregnancy causes placental defects and deficits in oxygen delivery to the fetal organs under hypoxic stress. Dev Biol 422:171-185 (2017). PubMed: 27940158
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

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

    Poor
    Abreviews
    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

    Ikki Mitsui

    Verified customer

    Submitted Aug 28 2020

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

    Good
    Abreviews
    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

    Abcam user community

    Verified customer

    Submitted Jul 27 2018

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

    Good
    Abreviews
    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

    Dr. Varkha Agrawal

    Verified customer

    Submitted Jul 03 2013

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Mar 18 2013

    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

    Abcam Scientific Support

    Answered on Mar 18 2013

    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

    Abcam community

    Verified customer

    Asked on Sep 13 2012

    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

    Abcam Scientific Support

    Answered on Sep 13 2012

    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

    Abcam community

    Verified customer

    Asked on Sep 12 2012

    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

    Abcam Scientific Support

    Answered on Sep 12 2012

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Sep 11 2012

    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

    Abcam Scientific Support

    Answered on Sep 11 2012

    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

    Abcam community

    Verified customer

    Asked on Jan 13 2012

    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

    Abcam Scientific Support

    Answered on Jan 13 2012

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

    Excellent
    Abreviews
    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

    Abcam user community

    Verified customer

    Submitted Jan 28 2009

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