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Assay Kits & ELISAs

Green Fluorescent FAM-Phe-CMK Serine Protease Assay Kit

Product Sizes
25 tests
£274.00
945-25TESTS
About this Product
SKU:
945
Application:
Flow Cytometry
CE/IVD:
RUO
Extra Details:
Quantify intracellular active chymotrypsin-like serine protease activity in vitro. The CMK group alkylates the active site histidine residue. Analyze your samples using a flow cytometer, fluorescence microscope, or fluorescence plate reader., ICT
Immunogen:
Chymotrypsin-like enzymes
Physical State:
Cell Culture, Tissue
Purification:
Intracellular serine protease activity was monitored over time in Jurkat cells exposed to 1 uM staurosporine. Cells were treated with staurosporine for 1, 2, 3, and 4 hours to induce apoptosis and inc
Shipping Conditions:
Blue Ice
Storage Conditions:
2-8[o]C, -20[o]C
Type:
Assay Kits: Detection
Figure 1. Jurkat cells were treated with staurosporine for 1, 2, 3, and 4 hours to induce apoptosis and increase serine protease activity (green), or were untreated (gray). Samples were stained with FAM-Phe-CMK (Kit 945/946) for 1 hour at 37°C, washed, and then analyzed using a flow cytometer. The amount of serine protease activity detected positively correlated to the duration of the exposure period. Data courtesy of Dr. Kristi Strandberg (ICT 231:75-79).
Figure 2. Jurkat cells were exposed to staurosporine to induce apoptosis and increase serine protease activity (upper), or left untreated (lower), then stained with FAM-Phe-CMK (Cat. 945/946), 660-VAD-FMK (cat. 9120), and Hoechst 33342. Imaged using EGFP (Ex 470/30, Em 530/50), Cy5 (Ex 620/60, Em 700/75), and DAPI (Ex 375/28, Em 460/50) LED filter cubes at 20X. Staurosporine increased the intracellular levels of both serine proteases and caspases. Data courtesy of Dr. K. Strandberg (ICT 231:73).
Figure 3. Jurkat cells were treated with a non-stimulating control or with an agent to stimulate serine protease activity. Cells were then labeled with FAM-Phe-CMK (Kit 945/946). There is a significant difference in relative fluorescence units (RFU) between the 2 experiments, but the ratio of non-stimulated:stimulated RFU was almost the same in each case (1:3.6 in experiment 1, and 1:3.9 in experiment 2). Data courtesy of Ms. Tracy Hanson (ICT 200:73).
Figure 4. FAM-Phe-CMK (Kit 945/946) was used to inhibit the enzyme Zmpste24, which cleaves prelamin A to generates a 31 kDa product. When Zmpste24 was inhibited by FAM-Phe-CMK (FFCK) much less 31 kDa product was detected. The zinc metalloprotease inhibitor 1,10-orthophenanthroline (Opa EDTA) has little effect on the cleavage of prelamin. Data courtesy of D. P. Corrigan, East Tennessee State University, Johnson City, TN.
Figure 5. HL-60 cells were treated with FAM-Phe-CMK (Kit 945/946) for 1 hour, then with staurosporine for an additional 2 hours. Cells treated with staurosporine had a higher level of green fluorescence (B) than untreated cells (A). Western blot of cell lysates with an anti-fluorescein antibody indicated a 62 kDa protein (C). Data courtesy of Dr. Catherine Stenson- Cox, National University of Ireland, Galway.