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Silver Staining Protocol for Minigels

Author: Karen Betancourt Moreira, updated date: , view: 224, Q&A: 0
Tags: Protein and Silver Stain


Approximately 100 mL of each solution are required per gel.  Add the formaldehyde just prior to use and have two liters of distilled water ready for rinsing.

  1. Fix gels 1hr to overnight in 50% methanol/12 % acetic acid/0.05% formaldehyde.

  2. Wash gels 2 X 10 min. in 50% ethanol.

  3. Pretreat for 1 min. with Na2S2O3 (100X stock is 2% w/v).

  4. Rinse 4 X 20 s with water.

  5. Incubate for 20 min with 2g/liter AgNO3 and 0.75 ml/L formaldehyde made fresh.  Do not pour silver down the sink!

  6. Rinse 4 X 20 s with water.

  7. Develop 10 min in 30 g/liter Na2CO3, 0.5 ml/liter formaldehyde, 200 microliters/liter Na2S2O3. (variable time)

  8. Rinse in distilled water.

  9. Stop reaction in 50% methanol, 12% acetic acid. 10 min

  10. Dry and analyze gels.

  1. 50 mls Methanol

12 mls Acetic Acid

50 ul Formaldehyde

38 mls water

     3)  50 mg Na2S2O3 in 1.5 ml water, add 1ml to 100 ml water

     5) to 100 mls of water add:

0.2 g AgNO3

75 ul formaldehyde

     7) to 100 mls of water add:

3g Na2CO3

50 ul formaldehyde

20 ul 100X stock

     9) 50 ml Methanol

12 ml Acetic acid

38 ml water

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