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).
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).
anti-Perforin (human), mAb (deltaG9) (APC)
ANC-358-060
Product group Antibodies
Overview
- SupplierAncell Corporation
- Product Nameanti-Perforin (human), mAb (deltaG9) (APC)
- Delivery Days Customer10
- CertificationResearch Use Only
- ClonalityMonoclonal
- Scientific DescriptionHuman 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. Antibody colne deltaG9 recognizes the human and rat perforin molecule. It has been reported to bind to pre-formed perforin granules, but not granule-independent perforin. Isotype: Mouse IgG2bkappa. Immunogen: Purified (With Antibiotics) granules from human YT lymphoma cell line. Applications: FACS, IHC. 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.
- Storage Instruction2°C to 8°C
- UNSPSC12352203