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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
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.
CAS Number	n/a
Clone Type	Monoclonal
Isotype	IgG2b,k
Clone No	7H179
Host	Mouse
Source	Human
Concentration	~1mg/ml
Form	Supplied as a liquid in PBS, pH 7.5, 0.2% BSA, 0.04% sodium azide, 5% glycerol. Labeled with biotin.
Purity	Purified by Protein A affinity chromatography.
Immunogen	Purified (With Antibiotics) granules from human YT lymphoma cell line
Specificity	Recognizes human perforin. Mr~ 70kD.
Important Note	This 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.