Apoptosis Assays and Reagents


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 OncoImmunin, Inc. has designed, synthesized, validated, and patented a new class of cell permeable fluorogenic protease substrates. Unique aspects of these probes are: (1) their ability to cross intact cell membranes enables measurement of intracellular protease activities in live cells, (2) incorporation of amino acid sequence information from both sides of protease cleavage sites. i.e. P and P' residues, up to a total of 10 amino acid residues, provides complete protease recognition sequences,  (3) conformation of substrates' peptide moieties assume loop or extended beta sheet conformations resulting in substrates retaining protease in vivo specificity and  (4) the choice of fluorophores  permits simultaneous measurement of multiple proteolytic activities in biologic samples.


PhiPhiLux® is OncoImmunin, Inc.'s novel and unique class of fluorogenic apoptosis assay substrates for the detection and measurement of caspase 3 and caspase 3-like activities in living cells (for other apoptosis-specific fluorogenic substrates [CaspaLux® family], see below). The presence of the amino acid sequence of DEVDGI allows one to determine the apoptosis specific, intracellular caspase-3 (CPP32) and caspase-3-like activities by flow cytometry or fluorescence microscopy. By judicious choice of fluorophores, either green or red fluorescence (or under license, far red) may be observed. Cells are neither permeabilized nor fixed. The principle used in the design of these apoptosis reagents was derived from exciton theory.

See the PhiPhiLux® review which appeared in The Scientist


Detailed PhiPhiLux® instructions are available:
(click icon for download):
onc1_gr.gif (743 bytes)PhiPhiLux® G1D2 green fluorescence
onc1_gr.gif (743 bytes)PhiPhiLux® G2D2  red fluorescence

Data from apoptosis studies


Treatment of a breast tumor cell (231-01) line with TRAIL:
Activation of caspase 8 (red = cleavage of CaspaLux® 8)


No other protease assay is like it!

Specific- peptide sequence is selected to match the desired protease
Sensitive- this is fluorescence, after all!
Broadly applicable - works not only for clear solutions but also cell suspensions

Give PhiPhiLux a try!

A PhiPhiLux® kit can be purchased using a Visa/MasterCard credit card by phone, FAX, or e-mail.
Call for special introductory prices.
PhiPhiLux® is US patented; additional US patent and PCT applications are pending.

With OncoImmunin's CaspaLux ® substrates, the activities of caspases-1,6,8 and -9 can be measured. Moreover, by combining the 5 different substrates in complementary colors, multiple caspase activity can be determined simultaneously in live cells by both flow cytometry and microscopy.  A high throughput assay is also available. 

PhiPhiLux cell-permeable fluorogenic substrates for caspases are available in two colors:
(click on icon to download protocol)
Caspase green red
Caspase 1 onc1_gr.gif (743 bytes)CaspaLux®-1 - E1D2 onc1_gr.gif (743 bytes)CaspaLux®-1 - E2D2
Caspase 6 onc1_gr.gif (743 bytes)CaspaLux®-6 - J1D2 onc1_gr.gif (743 bytes)CaspaLux®-6 - J2D2
Caspase 8 onc1_gr.gif (743 bytes)CaspaLux®-8 - L1D2 onc1_gr.gif (743 bytes)CaspaLux®-8 - L2D2
Caspase 9 onc1_gr.gif (743 bytes)CaspaLux®-9 - M1D2 onc1_gr.gif (743 bytes)CaspaLux®-9 - M2D2


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OncoImmunin, Inc.,
207A Perry Parkway, Suite 6
Gaithersburg, MD 20877
Phone: (301) 987-7881
FAX: (301) 987-7882





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