DAz-1

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
This experimental protocol is designed to identify protein oxidation in living cells.
1. Cell labeling (live cells): 1.1 Incubate the cells (such as Jurkat cells) with DAz-1 (1-10 mM) in a culture medium containing 2% FBS at 37°C for 1-2 hours.
1.2 Wash the cells three times with PBS to remove the excess probes.
1.3 Lysis: Prepare the cell lysis buffer (for example, using RIPA or an equivalent lysis solution).
2. Bioorthogonal Labeling : 2.1 Incubate the lysate with 100 μM p-biotin and 5 mM DTT at 37°C for 2 hours.
2.2 Terminate the reaction by precipitating with acetone (1 mL of cold acetone, incubated at -80°C for 1 hour, then centrifuged at 20,000 g for 20 minutes).
2.3 Resuspend the precipitate in the SDS-PAGE sample buffer.
3. Detection: Analyzed using streptavidin blotting (Western Blot).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.