Lactose Dehydrogenase (LDH) and PrestoBlue (PB) assay are biochemical assays to assess the cytotoxicity and viability of the cells in cell culture experiment respectively
Material Name | Quantity | Company | Catalog Number | Comment | |
---|---|---|---|---|---|
96-wells plate | Depends on the number of wells to be seeded (1 per 96 wells) | to contain the seeded cells | |||
1% serum medium | Depends on the number of wells to be seeded and their size | to support the seeded cells to wash the seeded cells | |||
0.25% trypsin | 5ml for each flask of cultured cells | to detach the cells from flask | |||
full culture medium | 5ml for each flask (for termination of trypsin effect) + 15ml for each subculture | to stop trypsinization and to subculture the cells | |||
hemacytometer | 1 | to quantify the cells | |||
multi-pipette (optional) | 1 | to seed the cells | |||
DPBS | 10ml for each flask of cultured cells | to wash the culture | |||
centrifugator | 1 | for centrifugation | |||
microscope | 1 | for cell quantification | |||
compound of interest | Depends on the experimental design | to stimulate the cells | |||
1 | to pipette out the supernatant to pipette out the supernatant and load the LDH reagent | ||||
LDH assay reagent | 1/2 of the supernatant (1% serum medium) used to seed the cells | to assess the released LDH | |||
lysis buffer | 10ul for each well in the Maximum control set | to induce the maximum LDH release | |||
PB assay reagent (10X) | 1/10 of the supernatant (1% serum medium) used to seed the cells | to assess the cell viability | |||
serum free medium | 2/5 of the supernatant (1% serum medium) used to seed the cells | to dilute the PB assay reagent | |||
plastic container | 2 | to contain the LDH and PB reagent | |||
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