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anti-Perforin (human), mAb (deltaG9) (Biotin)

Research Use Only
ANC-358-030
Ancell Corporation
ApplicationsFlow Cytometry, ImmunoHistoChemistry
Product group Antibodies
ReactivityHuman, Primate
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Overview

  • Supplier
    Ancell Corporation
  • Product Name
    anti-Perforin (human), mAb (deltaG9) (Biotin)
  • Delivery Days Customer
    10
  • Applications
    Flow Cytometry, ImmunoHistoChemistry
  • Certification
    Research Use Only
  • Clonality
    Monoclonal
  • Clone ID
    deltaG9
  • Conjugate
    Biotin
  • Formulation
    Liquid
  • Host
    Mouse
  • Isotype
    IgG2b
  • Scientific Description
    Human perforin (cytolysin) is believed to be one of the major effector molecules used by cytotoxic T cells and NK cells to mediate targeted cell lysis. Perforin expression is constitutive on NK cells, but increases in resting CD8+ cytotoxic cells upon activation. - Monoclonal Antibody. Isotype: Mouse IgG2bkappa. Clone: deltaG9. Applications: FACS, IHC. 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl, 5% Glycerol, 0.2% BSA, 0.04% NaN3 (as a preservative). Human perforin (cytolysin) is believed to be one of the major effector molecules used by cytotoxic T cells and NK cells to mediate targeted cell lysis. Perforin expression is constitutive on NK cells, but increases in resting CD8+ cytotoxic cells upon activation.
  • Reactivity
    Human, Primate
  • Storage Instruction
    2°C to 8°C
  • UNSPSC
    12352203

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Binding of anti-Perforin/APC to fixed permeabolized stimulated human PBL. Methods: Ficoll-prepared human peripheral blood mononuclear cells were stimulated by incubating 2 days at 5 x 10^6 cells/ml in RPMI 10% FBS media including 5microg/ml Phytohemagglutinin-P. Cells were then fixed for 30 min with 2% buffered formaldehyde after which they were washed two times. Subsequent reagent incubations and washes were done using a buffer containing 0.3% Saponin to permeabolize cells. Five x 10^5 cells per tube were washed and incubated 45 min on ice with 80microl of anti-Perforin/APC at a 1:50 dilution (5microg/ml). Cells were washed three times, fixed and analyzed by FACS. A net 16.1% subpopulation of the cells shifted positively with a mean shift of 1.49 log10 fluorescent units when compared to an IgG2b/R-PE isotype Control (#ANC-284-060). Binding was blocked when cells were pre-incubated 10 min with 20microl of 0.5mg/ml unlabeled anti-perforin antibody (#ANC-358-020).
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Binding of anti-Perforin/APC to fixed permeabolized stimulated human PBL. Methods: Ficoll-prepared human peripheral blood mononuclear cells were stimulated by incubating 2 days at 5 x 10^6 cells/ml in RPMI 10% FBS media including 5microg/ml Phytohemagglutinin-P. Cells were then fixed for 30 min with 2% buffered formaldehyde after which they were washed two times. Subsequent reagent incubations and washes were done using a buffer containing 0.3% Saponin to permeabolize cells. Five x 10^5 cells per tube were washed and incubated 45 min on ice with 80microl of anti-Perforin/APC at a 1:50 dilution (5microg/ml). Cells were washed three times, fixed and analyzed by FACS. A net 16.1% subpopulation of the cells shifted positively with a mean shift of 1.49 log10 fluorescent units when compared to an IgG2b/R-PE isotype Control (#ANC-284-060). Binding was blocked when cells were pre-incubated 10 min with 20microl of 0.5mg/ml unlabeled anti-perforin antibody (#ANC-358-020).
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