Protein concentration Amido black assay

material: Amido black solution: 10% (v/v)Acectic acid, 90% (v/v)MeOH, 0.05% (v/v) Amido black; Destain: 90% (v/v)EtoH; 10% (v/v) Acetic acid …
duration: 1h

• transfer 20ul of protein extract into new Eppendorf tubes; pipet samples for the calibration curve (see table 1)
• add 1 ml amido black solution; vortex
• centrifuge for 15 min at max. speed, RT
• discard supernatant
• add 1 ml destain solution
• mix by inversion
• centrifuge for 15 min at max. speed, RT
• discard supernatant
• air-dry the pellet (takes about 15 min)
• add 1ml 0.2 M NaOH; vortex
• transfer the samplkes to disposable cuvettes
• measure the OD620
• calculate the concentration of your protein samples