BODIPY FL SSE

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
BODIPY FL SSE assay (to quantify GAPDH)^[1]:
1.Make a 2 mg/ mL stock of GAPDH (MW 37 kDa) in PBS (pH 7.4).
2.Add 54 nM of BODIPY-FL-SSE to 1 mL of the 2 mg/ mL (54 μM) GAPDH solution. Incubate the mixture for 4 h on ice.
3.Dialyze the reaction mixture against 500 ml of PBS, pH 7.4, in a 10,000 MW-cutoff dialysis cassette for 1 h at 4 °C. Change the buffer and dialyze again overnight. Change the dialysis buffer once more the following morning and dialyze further for 1 h.
4.Run the dialyzed sample through a Zeba desalting column at 1000 g for 2 min.
5.Analyze the BD-GAPDH spectrophotometrically in a quartz cuvette. Set the wavelength scan to 250-700 nm. Blank the instrument against PBS and add the BD-GAPDH to the cuvette.

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