EDC Impressum & DSE

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!

Mai 30, 2018

ProQNewRecipeAS

Chr. Hentschker, Mikrobiologie Greifswald (Germany)

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

Pro-Q Diamond Stain

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