EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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EDC’s SDS & 2D Silver Staining Protocol

Ag-staining recipe for EDC’s ElphoGels SDS
This special SDS staining recipe is necessary because EDC’s SDSGels use different buffer molecules to store and run them.

For 2D-gels this staining should be mass-spectrometer compatible:
No Glutardialdehyde in the Conditioner!

September 25, 2018

steps

reagent 

volume

time
(0.4-0.6 mm)

1 Fixing

 40% (v/v) Ethanol / 10% (v/v) Acetic Acid, 1% Citric Acid
(80 ml EtOH / 20 ml HAc + 2 g Citric Acid in 200 ml)

1 x 200 ml

60 min

2 Washing

H2O dist

4 x 200 ml

4 x 10 min

3 Conditioning

30% (v/v) Ethanol; 0,325 % (w/v) Sodium Thiosulphate
165 mM Sodium Acetate; 2.5 % Glutardialdehyde (w/v), 0.07% (w/v) Benzenesulfonic acid
(60 ml EtOH / 650 mg Thiosulfate / 4.5 g NaAc / 20 ml Benzenesulfonic stock sol., 20 ml Glutardialdehyde (25%) in 200ml)

200 ml

30 min

4 Washing

H2O dist (place gel on the glas tray bottom surface side up)

4 x 200 ml

4 x 5 min

5 Silvering

0.2 % (w/v) AgNO3 /0.04% (v/v) Formaldehyde (37%), 0.07% Benzenesulfonic Acid
8 ml AgNO3 - stock sol. / 20 ml Benzenesulfonic stock sol./ 260 µl Formaldehyde in 200 ml)

200 ml

45 min

6 Washing

H2O dist

3 x 200 ml

3 x 1 min

7 Developing

2.5 % Na2CO3 (w/v) 0.05% (w/v) Formaldehyde / 0.002% (w/v) Sodium-Thiosulfate
(5 g NaCO3 / 260 ul Formaldehyde (37%) / 200 µl Sodium-Thiosulfate (2%, fresh!) in 200 ml)

1 x 200 ml

2 - 4 min

8 Stopping/
Preserving

10% (v/v) HAc, 10% (v/v) Glycerol
(20 ml HAc / 20 ml Glycerol in 200 ml)

200 ml

20 min

9 Drying

air dry , on the support-film
then roll on a polyester-sheet

---

overnight

droptest

Droplet-test: within 3 seconds

Stock Solutions
1.  Benzenesulfonic (Merck 8.00468.0250), 0.7 % (10-fold solution) 1,4 g / 200 ml
2.  AgNO3: 5 % (20-fold solution), 5 g in 100 ml H2O dist.
3.  Na2CO3: 25% (10-fold) solution, 25 g (water-free!) in 100 ml H2O dist.
4.  Thiosulfate: 2%: 200 mg / 10 ml (prepare daily fresh!)
AgNO3, Na2CO3 (water-free), Sodiumthiosulfate are from Merck
Benzenesulfonic acid, Ethanol, Acetic Acid, Formaldehyd (37%), Glutardialdehyde, Citric Acid and Glycerol are from Merck Darmstadt (Germany)

Abbr.: HAc = Acetic Acid, NaAc = Sodium Acetate

ElphoGel SDS 10% 25S stined with the recipe above

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