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Assay Kits & ELISAs

Pyroptosis/Caspase-1 Assay - Far Red Fluorescent

Product Sizes
25 tests
£455.00
9158-25TESTS
About this Product
SKU:
9158
Application:
Flow Cytometry
CE/IVD:
RUO
Extra Details:
This Pyroptosis/Caspase-1 Assay Kit utilizes our FLICA® technology to assess pyroptotic cells by detecting caspase-1 activation. Analyze the far-red fluorescent signal using flow cytometry or fluorescence microscope., ICT
Immunogen:
Caspase 1
Physical State:
Cell Culture
Purification:
Caspase-1 activity in nigericin-treated Jurkat cells. Jurkat cells were mock treated with a negative control (Non-Induced, Panels A-C), or Nigericin (10 uM) to induce NLRP3 inflammasome and caspase-1
Shipping Conditions:
Blue Ice
Storage Conditions:
2-8[o]C
Type:
Assay Kits: Cell Viability
Figure 1. Signals leading to activation of the NLRP3 inflammasome.
Figure 2. THP-1 cells were treated with PMA (5 ng/mL) followed by LPS (100 ng/mL for 1 hour, lower row of images), or were untreated (upper row of images), and were then immediately stained with 660-YVAD-FMK for 1 hour at 37°C (concurrent with treatment). Cells were then stained with Hoechst 33342 and imaged. Intracellular caspase-1 activity was detected using Cy5 (Ex 620/60, Em 700/75) and DAPI (Ex 375/28, Em 460/50) LED filter cubes at 20X. Data courtesy of Mrs. Tracy Murphy (ICT 230:32).
Figure 3. THP-1 cells were spiked with 660-YVAD-FMK and then treated either with LPS (1 ug/mL for 3 hours), Nigericin (20 uM for 30 minutes), or LPS + Nigericin (1 ug/mL LPS for 3 hours, followed by 20 uM Nigericin for 30 minutes). Cells were analyzed by flow cytometer. LPS or Nigericin increased caspase-1 activity approx. 4-fold compared to the negative control. LPS + Nigericin increased caspase-1 activity almost 2-fold over either treatment alone. Data courtesy of Mrs. T. Murphy, ICT (220:95).
Figure 4. Jurkat cells were treated with a negative control (Panels A-C), or nigericin (10 uM) to induce NLRP3 inflammasome and caspase-1 activation (Panels D-F). Samples were stained with 660-YVAD-FMK, followed by Hoechst 33342 (to label nuclei). Significant caspase-1 activity is seen only after nigericin treatment (D). Panels C & F are overlays of the blue fluorescence, red fluorescence, and DIC channels into a single combined image. Data courtesy of Mrs. Tracy Murphy, ICT (220:98).
Figure 5. A common pool of Jurkat cells was spiked with 660-YVAD-FMK and divided into separate treatment groups. Samples were treated with 1.25, 2.5, 5, 10, and 20 uM Nigericin and incubated at 37°C for 0, 12, 24, 36, and 48 hours. Cells were then washed and analyzed by flow cytometer. Caspase-1 activity correlated with both the duration of exposure and the nigericin concentration. Data courtesy of Mrs. Tracy Murphy, ICT (230:01).