P3340-15B

Perforin (Cytolysin) (Biotin)

Size

£639.00

SKU:: P3340-15B
Purity:: Purified by Protein A affinity chromatography.
Storage Conditions:: 4°C (-20°C Glycerol)
Supplier:: United States Biological
Host:: Mouse
Immunogen:: Purified (With Antibiotics) granules from human YT lymphoma cell line
Clone:: 7H179
Format:: Supplied as a liquid in PBS, pH 7.5, 0.2% BSA, 0.04% sodium azide, 5% glycerol. Labeled with biotin.
Extra Details:: 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. Applications: Suitable for use in Flow Cytometry and Immunohistochemistry. Other applications not tested. Recommended Dilution: Flow Cytometry: 2ug/ml. Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. 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 Stability: May 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.

Shipping Conditions:: Blue Ice