P3340-15A

Perforin (Cytolysin)

Size

£557.00

SKU:: P3340-15A
Purity:: Purified by Protein A affinity chromatography from tissue culture supernatant.
Storage Conditions:: -20°C
Supplier:: United States Biological
Host:: Mouse
Immunogen:: Purified granules from human YT lymphoma cell line
Clone:: 7H179
Format:: Supplied as a liquid in FBS, 50mM sodium phosphate pH 7.5, 100mM potassium chloride, 150mM sodium chloride, 0.5 mg/ml gentamicin sulfate.
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: Immunohistochemistry: Frozen, Paraffin Optimal dilutions to be determined by the researcher. Protocol: 1. Ficoll prepared human peripheral white blood cells were stimulated by incubating 2 days at 5x10e6 cells/ml in RPMI 10% FBS media including 5 mg/ml Phytohemagglutinin-P. 2. Cells were then fixed for 30 minutes with 2% buffered formaldehyde after which they were washed two times. Note: Subsequent reagent incubations and washes were done using a buffer containing 0.3% Saponin to permeabolize cells. 3. Five x 10e5 cells per tube were washed and incubated 45 minutes on ice with 80ul of P3340-15A at 5mg/ml. 4. They were then washed twice and incubated with IgG (FITC) Goat anti-Mouse, after which they were washed three times, fixed and analyzed using a BD FACstar plus. 5. A net 32% population of the cells stained positive with a mean shift of 1.51 log10 fluorescent units when compared to a Mouse IgG2b negative control at a similar concentration. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Shipping Conditions:: Blue Ice