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Assay Kits & ELISAs

Pyroptosis/Caspase-1 Assay - Green Fluorescent

Product Sizes
25-50 tests
9145-25-50TESTS
About this Product
SKU:
9145
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 green fluorescent signal using fluorescence microscopy, a fluorescent plate reader, or by flow cytometry., ImmunoChemistry Technologies
Immunogen:
Caspase 1
Physical State:
Cell Culture, Tissue
Purification:
THP-1 suspension cells were treated with PMA (5 ng/mL) to induce differentiation into macrophages. After 48 hours, PMA was removed and replaced with fresh medium that was spiked with LPS (10 ng/mL) to
Shipping Conditions:
Blue Ice
Storage Conditions:
2-8[o]C, -20[o]C
Type:
Assay Kits: Cell Viability
Figure 1. Signals leading to activation of the NLRP3 inflammasome.
Figure 2. C2BBe1 cells were infected with wild type Salmonella constitutively expressing mCherry (red). After 9 hours, live cells were incubated with FAM-YVAD-FMK (green cell) for 1 hour. The infected cell (red) is undergoing pyroptosis, and shows increased green fluorescence compared to uninfected cells due to caspase-1 activation. Data courtesy Knodler, Leigh A., et al. Dissemination of invasive Salmonella via bacterial-induced extrusion of mucosal epithelia. PNAS. 107:41, 17733-17738 (2010).
Figure 3. THP-1 cells were treated with either a negative control or PMA (5 ng/mL) followed by LPS (10 ng/mL) to induce differentiation and pyroptosis. Cells were stained with FAM-YVAD-FMK and analyzed using a fluorescence plate reader (488 nm excitation and 530 nm emission using a 515 nm cut-off filter). In the pyroptotic (induced) cells, the green fluorescent signal was nearly 2-fold greater than in the negative (non-induced) cells. Data courtesy of Mrs. Tracy Murphy, ICT (207:13-14).
Figure 4. THP-1 cells were treated with either a negative control (A), or PMA (5 ng/mL) followed by LPS (10 ng/mL) to induce differentiation and caspase-1 activation. Cells were stained with FAM-YVAD-FMK and analyzed by photomicroscope using phase contrast and fluorescence optics. Many more bright green cells are seen in the induced sample, indicating increased caspase-1 activity (B), as compared to the uninduced sample (A). Data courtesy of Dr. Brian Lee, ICT (207:11).
Figure 5. A common pool of Jurkat cells was spiked with FAM-YVAD-FMK and then divided into separate treatment groups. Cells were treated with 10 uM nigericin at 37°C for 1, 3, 6, 12, and 24 hours. Following treatment, the cells were washed and analyzed by flow cytometer. The longer the cells were exposed to nigericin, the greater the proportion of caspase-1 positive cells found in the sample. Data courtesy of Mrs. Tracy Murphy, ICT (220:78).