Hello Shaiza Talib,

Different cell types exhibit varying sensitivities to hydrogen peroxide-induced oxidative stress. Some cells might be more susceptible to damage than others, requiring shorter or longer exposure times to achieve similar effects.

The extent of cell detachment depends on both the concentration of hydrogen peroxide used and the duration of exposure. Hydrogen peroxide can induce cell death pathways like apoptosis and necrosis, which can lead to cell detachment. It can also affect the cell's cytoskeleton and adhesion molecules, causing them to lose their grip on the culture surface. 

You may have to optimize both the concentration of hydrogen peroxide and the time of exposure. You may start with a range of concentrations, such as 10µM, 20µM, 50µM, 80µM and 100µM. Also, use different time points such as 10mins, 30mins, 60mins, 90mins and 120mins. For instance, make an experimental set of 10mins having five different concentrations. In this way, you will have 5 different sets with 5 different concentrations. From these you may choose the most appropriate hydrogen peroxide concentration and time of exposure.

Best,