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Assay Kits & ELISAs

Red Fluorescent SR-FLICA[R] Poly (active) Caspase (VAD) Assay Kit

Product Sizes
25 tests
916-25TESTS
About this Product
SKU:
916
Application:
Flow Cytometry
CE/IVD:
RUO
Extra Details:
This in vitro assay employs the red fluorescent inhibitor probe SR-VAD-FMK to label active caspase enzymes in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry., ImmunoChemistry Technologies
Immunogen:
Poly caspases
Physical State:
Cell Culture
Purification:
Jurkat cells were treated with 1 uM staurosporine to induce caspase activity (top), or a negative control (bottom) for 3 hours, incubated with ICT's red poly caspases inhibitor probe, SR-VAD-FMK, for
Shipping Conditions:
Blue Ice
Storage Conditions:
2-8[o]C
Type:
Assay Kits: Cell Viability
Figure 1. HL-60 cells were treated with camptothecin (cat. 6210), followed by SR-VAD-FMK (cat. 916/917) and green FAM-FLISP serine protease inhibitor FFCK (cat. 946), and then analyzed on a scanning laser cytometer. Active caspases stain red (X-axis) and active serine proteases stain green (Y-axis). Co-localization of caspase and serine protease activity is evident in B, C, D, and G. Data courtesy of Dr. J. Grabarek, Brander Cancer Center, NY, and Pomeranian School of Medicine, Szczecin, Poland.
Figure 2. Paramecium were starved for 12-24 hours in 5 mM KCl to induce autogamy. SR-VAD-FMK (cat. 916/917), was added to the live Paramecium for 30 minutes in the dark. Cells were then washed and Hoechst 33342 was added. Cells were incubated in the dark for 10 minutes, washed, and applied to coverslips. Images were captured using a Nikon TE-300 Epi-fluorescent microscope with a SPOT-RT digital camera and merged using SPOT software. Data courtesy of Dr. W. E. Bell, Virginia Military Institute.
Figure 3. Jurkat cells and HL-60 cells were treated with either DMSO (negative) or staurosporine (apoptotic) for 3 hours at 37°C, and then labeled with SR-VAD-FMK (cat. 916/917) for 60 minutes at 37°C. Cells were read on a fluorescence plate reader set at 550 nm excitation and 595 nm emission with a 570 nm cut-off filter. Staurosporine increased red fluorescence 15X in the Jurkat cells (1.9 to 29.8) but only 5X in HL-60 cells (3.0 to 14.7). Data courtesy of Dr. Brian W. Lee, ICT.
Figure 4. K562 cells were stained with SR-VAD-FMK, followed by 7-AAD red live/dead stain (cat. 6163), and analyzed using bi-color flow cytometry. Unstained live cells have minimal fluorescence (67%, lower left); early apoptotic cells fluoresce orangy-red (8%; lower right); late apoptotic cells are dually stained with SR-FLICA and 7-AAD (23%; upper right); necrotic cells fluoresce red with 7-AAD (2%; upper left). Data courtesy of Dr. Michael Olin, University of Minnesota.
Figure 5. K562 cells were treated with staurosporine (cat. 6212) to induce apoptosis and potentially late apoptosis-associated necrosis, or DMSO. Cells were stained with SR-VAD-FMK, followed by 7-AAD and analyzed on a flow cytometer. Apoptotic cells fluoresce orangy-red with SR-FLICA and shift to the right on the X-axis. The treatment did not significantly increase necrosis in the K562 cell population but it did increase apoptosis. Data courtesy of Dr. Michael Olin, University of Minnesota.