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Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Fluorescence Visualization of Phosphorylated Proteins


	Use Invitrogen’s Pro-Q manual: 1. Protocol: take the row “Standart-Protocol” for “BisTris-Gels”. 2. Step 1-3: Do not take 50% MeOH but 40% (50% is too much for the support-film!) Ethanol is also working!


	Pro-Q-Diamond protocol (slightly changed) for EDC’s 2d-gels (C.Hentschker, Mikrobiologie Greifswald, Germany)

Step	Reagent	Protocol
Step 1: Fix 1	40% methanol (or ethanol), 10% acetic acid	100 mL, 30 - 60 min
Step 2: Fix 2	40% methanol (or ethanol), 10% acetic acid	100 mL, overnight
Step 3: Fix 3	40% methanol (or ethanol), 10% acetic acid	100 mL, 30-60 min
Step 4-6: Wash	Ultrapure water	100 mL, 15 min 3 times
Step 7: Stain	Pro-Q® Diamond stain	60 mL, 3 h
Step 8-10: Destain	Pro-Q® Diamond destain solution or: 20% acetonitrile 50 mM sodium acetate, pH 4	80 - 100 mL, 30 min 3 times
Step 11-12: Wash	Ultrapure water	100 mL, 30 min 5 times
Total solution changes	12	---
Total time	---	overnight plus ~8 hr


	light sensitive!

April 28, 2023


	Chr. Hentschker, Mikrobiologie Greifswald (Germany)


	Pro-Q-Staining (black <--> white) according the table above


	Chr. Hentschker, Mikrobiologie Greifswald (Germany)


	Green spots are all proteins by: Flamingo (Bio-Rad) --> see: Flamingo-Stain Red spots are the phosphorylated proteins Yellow spots are all proteins + phorphorylated proteins


	EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)