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You are here:Home » Antibodies » Abs to Apoptosis » Anti -Perforin (Cytolysin) (Biotin)

Anti -Perforin (Cytolysin) (Biotin)

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Specifications

Clone Host Grade Applications
Monoclonal Mouse Affinity Purified IH FC
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.
Catalog #P3340-15B
ApplicationsSuitable for use in Flow Cytometry and Immunohistochemistry. Other applications not tested.
Recommended DilutionFlow Cytometry: 2ug/ml.
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol1. Ficoll preparediscontinued human peripheral white blood cells were stimulated by incubating 1 day in culture at 5x10e6 cells/ml with 10ng/ml PMA with 2uM Ionomycin.
2. Harvest cell and fix for 30 min. with 2% buffered formaldehyde. Wash cells 2x with buffer.
3. Subsequent incubations and washes are carried out with buffer containing 0.3% Saponin to permeabilize cells.
4. Incubate 5x10e5 cells/tube for 45 min. on ice with 80ul of
P3340-15B at 2ug/ml.
5. Wash 2x and incubate with secondary reagent, Streptavidin (PE) (S7973-88P).
6. Wash cells 3x, fix and analyze by FACS.
7. A net 4% sub-population of cells stained positive with a mean shift of 1.1 log10 fluorescent units. Binding was blocked when cells were pre-incubated with 20ul of 0.5mg/ml of P3340-15B.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Clone TypeMonoclonal
IsotypeIgG2b,k
Clone No7H179
HostMouse
SourceHuman
Concentration~1mg/ml
FormSupplied as a liquid in PBS, pH 7.5, 0.2% BSA, 0.04% sodium azide, 5% glycerol. Labeled with biotin.
PurityPurified by Protein A affinity chromatography.
ImmunogenPurified (With Antibiotics) granules from human YT lymphoma cell line
SpecificityRecognizes human perforin. Mr~ 70kD.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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