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Caspase 1 fluorometric assay

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The Caspase-1 Fluorometric Assay is a laboratory tool designed to measure the activity of the caspase-1 enzyme. Caspase-1 is an important protease involved in various cellular processes, including inflammation and programmed cell death. The assay utilizes a fluorogenic substrate that emits a fluorescent signal upon cleavage by active caspase-1, allowing for the quantification of the enzyme's activity.

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Lab products found in correlation

Cytometric bead array, by BD (2 mentions) Mouse inflammation kit, by BD (2 mentions) Il β1 ready set go elisa, by Thermo Fisher Scientific (2 mentions)

2 protocols using caspase 1 fluorometric assay

1

Comprehensive Lung Inflammation Analysis

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To perform bronchoalveolar lavage (BAL), a 20-gauge angiocath ligated into the trachea and 1 ml of sterile PBS was instilled, then removed through the angiocath; the process was repeated three times. The samples were pooled and a 200 μl aliquot of the BAL fluid was placed in a cytospin and centrifuged at 500 g for 10 min. The pellet was re-suspended in 1 ml of PBS, plated on glass slides, stained with crystal violet and subjected to a blinded manual cell count and differential. Cytokine and chemokine levels in the supernatant were measured using the BD Cytometric Bead Array (BD Biosciences). Samples were analysed in triplicate using the Mouse Inflammation Kit (BD Biosciences) according to the instructions provided. This kit detects interleukin (IL)-6, MCP-1 and tumour necrosis factor (TNF)-α. Total transforming growth factor (TGF)-β1 was measured using the TGF-β1 RII DuoSet enzyme-linked immunosorbent assay (ELISA, R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. Total interleukin (IL)-1β was measured using the IL-β1 Ready-Set-GO ELISA (Ebioscience, San Diego, CA) according to the manufacturer’s instructions. Caspase-1 activity was measured using the Caspase-1 Fluorometric Assay (R&D Systems), according to the manufacturer’s instructions.
dos Santos G., Rogel M.R., Baker M.A., Troken J.R., Urich D., Morales-Nebreda L., Sennello J.A., Kutuzov M.A., Sitikov A., Davis J.M., Lam A.P., Cheresh P., Kamp D., Shumaker D.K., Budinger G.R, & Ridge K.M. (2015). Vimentin regulates activation of the NLRP3 inflammasome. Nature Communications, 6, 6574.
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2

Bronchoalveolar Lavage Immune Profiling

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To perform bronchoalveolar lavage (BAL), a 20-gauge angiocath ligated into the trachea and 1 ml of sterile PBS was instilled then removed through the angiocath; the process was repeated three times. The samples were pooled and a 200 μl aliquot of the BAL fluid was placed in a cytospin and centrifuged at 500 × g for 10 min. The pellet was re-suspended in 1 ml of PBS, plated on glass slides, stained with crystal violet and subjected to a blinded manual cell count and differential. Cytokine and chemokine levels in the supernatant were measured using the BD Cytometric Bead Array (BD Biosciences). Samples were analyzed in triplicate using the Mouse Inflammation Kit (BD Biosciences) according to the instructions provided. This kit detects interleukin (IL)-6, MCP-1, and tumor necrosis factor (TNF)-α. Total transforming growth factor (TGF)-β1 was measured using the TGF-β1 RII DuoSet ELISA (R&D Systems, Minneapolis, MN) according to the manufacturer's instructions. Total interleukin (IL)-1β was measured using the IL-β1 Ready-Set-GO ELISA (Ebioscience, San Diego, CA) according to the manufacturer's instructions. Caspase-1 activity was measured using the Caspase-1 Fluorometric Assay (R&D Systems), according to the manufacturer's instructions.
dos Santos G., Rogel M.R., Baker M.A., Troken J.R., Urich D., Morales-Nebreda L., Sennello J.A., Kutuzov M.A., Sitikov A., Davis J.M., Lam A.P., Cheresh P., Kamp D., Shumaker D.K., Budinger G.R, & Ridge K.M. (2015). Vimentin regulates activation of the NLRP3 inflammasome. Nature Communications, 6, 6574.
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