Rhodamine-N3 chloride

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
In-situ labeled sample processing, based on gel-based active protein analysis ^[2]. Let the cells be placed on ice to thaw, and add 200 μL of cold DPBS along with the protein inhibitor mixture to each sample.
2. Disrupt the cells and centrifuge at 100,000 × g for 45 minutes.
3. Take the supernatant (the soluble protein fraction), resuspend it in 200 μL of cold DPBS (with protease inhibitors).
4. Standardize the protein concentration of each sample to 1 mg/mL, with a volume of 43 μL.
5. Perform click chemical reactions on each sample using a solution with a final concentration of 25 μM Rhodamine-N3 chloride, 1 mM TCEP, 100 μM TBTA, and 1 mM CuSO4, with a final volume of 50 μL.
6. Incubate at room temperature for 1 hour, then add 20 μL of 4X SDS loading buffer.
7. Add the sample (30 μL) and perform electrophoresis on a 10% SDS-PAGE gel. Then, image it using a fluorescence scanner.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.