The activation of Caspase 1 in mouse brain samples were determined using Caspase 1 Colorimetric Assay Kit (YVAD, Merck Millipore, Burlington, MA) and following the manufacturer’s instructions. The assay determines the activity of Caspase 1 that recognizes the sequence YVAD. In brief, hippocampus and amygdale tissue samples from one brain hemisphere were homogenized in an ice-cold lysis buffer. The lysates were added to a Caspase 1 reaction buffer in a 96-well flat-bottom microplate. A substrate solution containing YVAD conjugated to chromophore p-nitroanilide (pNA) was added to each well and this was followed by incubation at 37 °C for 2 h. The quantification of Caspase 1 activity was carried out by the detection of pNA from the cleavage of the pNA-YVAD substrate by measuring light emission at 405 nm using a microplate reader (Molecular Devices, San Jose, CA).
Caspase 1 colorimetric assay kit
Manufactured by Merck Group
The Caspase 1 Colorimetric Assay Kit is a laboratory equipment product designed to detect and quantify the activity of Caspase 1, a key enzyme involved in the inflammatory response. The kit provides a colorimetric method for measuring Caspase 1 activity in cell or tissue lysates.
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2 protocols using caspase 1 colorimetric assay kit
1
Caspase 1 Activity Determination in Mouse Brain
2
Quantifying Caspase 1 Activity in Mouse Brain
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The activation of Caspase 1 in mouse brain samples were determined using Caspase 1 Colorimetric Assay Kit (YVAD, Merck Millipore, Burlington, MA) and following the manufacturer's instructions. The assay determines the activity of Caspase 1 that recognizes the sequence YVAD. In brief, hippocampus and amygdale tissue samples from one brain hemisphere were homogenized in an ice-cold lysis buffer. The lysates were added to a Caspase 1 reaction buffer in a 96-well flat-bottom microplate. A substrate solution containing YVAD conjugated to chromophore p-nitroanillide (pNA) was added to each well and this was followed by incubation at 37°C for 2 hours. The quantification of Caspase 1 activity was carried out by the detection of pNA from the cleavage of the pNA-YVAD substrate by measuring light emission at 405 nm using a microplate reader (Molecular Devices, San Jose, CA).
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