Technical Data
P3340-15B
Perforin (Cytolysin) (Biotin)
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.

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.
TypeIsotypeCloneGrade
MabIgG2b,k7H179Affinity Purified
SizeStorageShippingSourceHost
100ug4°C (-20°C Glycerol)Blue IceHumanMouse
Concentration:
~1mg/ml
Immunogen:
Purified (With Antibiotics) granules from human YT lymphoma cell line
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.5, 0.2% BSA, 0.04% sodium azide, 5% glycerol. Labeled with biotin.
Specificity:
Recognizes human perforin. Mr~ 70kD.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. M.J. Smyth, et al, (1990) J Exp Med 171: 1269-1281. 2. Hameed, A. et al, (1992) Am J Pathol 140: 1025-1030. 3. Jose, P.L. et al,  (1994) J Immunol 148: 3354-3360. 4. Schlesinger, B.C. & Cheng, L. (1994) Immunol 81: 291-295. 5. Liu, C.C. et al, (1995) Immunol Today 16: 194-201