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Assay Kits & ELISAs

Green Fluorescent Poly (active) Caspase FAM FLICA[R] Assay Kit (FAM-VAD-FMK)

Product Sizes
25 tests
91-25TESTS
About this Product
SKU:
91
Application:
Flow Cytometry
Extra Details:
This FLICA® (Fluorescent Labeled Inhibitors of CAspases) kit is designed to detect active caspase enzymes. This in vitro assay employs the fluorescent inhibitor probe FAM-VAD-FMK to label active caspase enzymes in living cells. Samples can be analyzed using fluorescence microscopy, a fluorescence plate reader, or by flow cytometry. Our Green Fluorescent Poly (active) Caspase FAM FLICA® Inhibitor Reagent (FAM-VAD-FMK) can be used as a general reagent to detect apoptosis as it is recognized by many types of activated caspases. The probe measures the intracellular process of apoptosis instead of a side-effect, such as the turnover of phosphatidyl serine, and eliminates the incidence of false positives that often plagues methods like Annexin V and TUNEL staining.
Immunogen:
Poly Caspases
Physical State:
Cell Culture
Shipping Conditions:
Blue Ice
Storage Conditions:
2-8[o]C
Type:
Assay Kits: Cell Viability
Figure 1. Fluorescent Microscopy Analysis of THP-1 Suspension Cells THP-1 cells were stained with Poly (active) Caspase FAM FLICA® Inhibitor Reagent (green), and Hoechst 33342 (Blue). Cells were incubated with staurosporine to induce apoptosis. Two photos were taken and superimposed. Only one cell of the three cells is apoptotic (middle) - it is stained positive for caspase activity with FAM-VAD-FMK. Data courtesy of Dr. Brian W. Lee.
Figure 2. Caspase Activity in Jurkat Cells Jurkat cells (T lymphocytes) were labeled with our Green Fluorescent Poly (active) Caspase FAM FLICA® Inhibitor Reagent (FAM-VAD-FMK) (catalog 92) and viewed under a fluorescence microscope. The grey DIC image (B) reveals five cells in the field of view, but only four of them fluoresce green (A). Four out of five cells are apoptotic and have active caspases present (green). Data courtesy of Dr. Brian W. Lee, ICT.
Figure 3. Single Color Analysis via Flow Cytometry Jurkat cells were treated with DMSO, a negative control (left), or staurosporine (right), then stained with our Green Fluorescent Poly (active) Caspase FAM FLICA® Inhibitor Reagent (catalog 92). The negative control exhibited caspase activity in only 3.3% of the cell population (P3, left histogram), whereas treatment with staurosporine induced caspase activity in 94.8% of the experimental cells (P3, right histogram). Courtesy of Mrs. Tracy Murphy.
Figure 4. Cells- a placebo (top), or apoptotic (bottom), stained with FAM-VAD-FMK and Propidium Iodide (PI). Caspase activity (green, FL-1) on the X-axis and necrosis (red, FL-2) on the Y-axis. (A) unstained live cells; (B) cells in early apoptosis fluoresce green with FAM-FLICA; (C) cells in late apoptosis are dually stained with FAM-FLICA and PI: they fluoresce green (active caspases) and red (the cell membrane has been permeabilized); and (D) necrotic cells fluoresce red. Courtesy of JH.
Figure 5. HL-60 cells treated with a drug, then stained with Green Fluorescent Poly (active) Caspase Inhibitor Reagent and Propidium Iodide (PI). Red X-axis (necrosis) and green on the Y-axis (apoptosis). (A) unstained live cells; (B) necrotic cells fluoresce red; (C) cells in late apoptosis stained with FAM-FLICA and PI: fluoresce green (active caspases) and red (cell membrane is compromised); and (D) cells in early apoptosis fluoresce green. Courtesy of Dr. Zbigniew Darzynkiewicz.