Mice were sacrificed by lethal i.p. injection of 500 mg/kg ketamine HCl (Ketathesia; Henry Schein Animal Health, Dublin, OH) and bled via the retro-orbital venous plexus. Blood was collected in BD Microtainer SST tubes (BD Biosciences, Franklin Lakes, NJ) containing polymer gel and 100 U heparin (Fresenius Kabi USA, Schaumburg, IL) at 0, 2, 4, and 6 h post-infection. Sera samples were purified, and alanine aminotransferase (ALT) concentration was determined by mixing 20 μL serum with 200 μL liquid ALT working buffer (Pointe Scientific, Canton, MI). The reaction was incubated for 180 s at 37°C. Absorbance at 340 nm was read at 0, 60, 120, and 180 s time points with a kinetic read function using a SpectraMax M2 Microplate Reader equipped with SoftMax Pro (v5.3b12) software (Molecular Devices, San Jose, CA). ΔOD340/min was used to calculate ALT concentration.
Softmax pro v5.3b12 software
Manufactured by Molecular Devices
SoftMax Pro (v5.3b12) software is a data acquisition and analysis software developed by Molecular Devices. It is designed to work with a range of Molecular Devices' microplate readers and spectrophotometers. The software enables users to collect, analyze, and manage data generated from these instruments.
Automatically generated - may contain errors
2 protocols using softmax pro v5.3b12 software
1
Kinetic ALT Assay for Murine Liver Injury
2
Kinetic ALT Assay for Murine Liver Injury
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Mice were sacrificed by lethal i.p. injection of 500 mg/kg ketamine HCl (Ketathesia; Henry Schein Animal Health, Dublin, OH) and bled via the retro-orbital venous plexus. Blood was collected in BD Microtainer SST tubes (BD Biosciences, Franklin Lakes, NJ) containing polymer gel and 100 U heparin (Fresenius Kabi USA, Schaumburg, IL) at 0, 2, 4, and 6 h post-infection. Sera samples were purified, and alanine aminotransferase (ALT) concentration was determined by mixing 20 μL serum with 200 μL liquid ALT working buffer (Pointe Scientific, Canton, MI). The reaction was incubated for 180 s at 37°C. Absorbance at 340 nm was read at 0, 60, 120, and 180 s time points with a kinetic read function using a SpectraMax M2 Microplate Reader equipped with SoftMax Pro (v5.3b12) software (Molecular Devices, San Jose, CA). ΔOD340/min was used to calculate ALT concentration.
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