Stock solutions

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Recipes for Stock Solutions

1. Ammonium Persulfate (10%)
   • 1 g container
   • 10 ml ddH2O
   • dissolve and divide into 1 ml aliquots
   • freeze at -20
2. Aprotinin (2 mg/ml)
   • 10 mg container
   • 5 ml ddH2O
   • dissolve into small vial
   • cover in alum. foil
   • store at 4 degrees
3. Coomassie Destaining Sol’n
   • 30 % methanol
   • 10 % acetic acid
   • ddH2O
4. Coomassie Staining Sol’n (can be reused)
   • 0.25 g coomassie Brilliant Blue R250
   • 90 ml methanol: H2O (1:1 v/v)
   • 10 ml acetic acid
5. EDTA (0.5 M) 500 ml sol’n
   • 73 g EDTA
   • ddH2O
   • Use ~35 g NaOH chips to dissolve all EDTA
   • pH 8.0
6. EGTA (0.5 M) 100 ml sol’n
   • 23.4 g EGTA
   • ddH2O
   • pH 8.0
7. HEPES (1 M) 100 ml sol’n
   • 24 g HEPES
   • ddH2O
   • pH 7.4
8. KCl 100 ml sol’n
   • 7.5 g KCl
   • ddH2O
9. Laemmli Buffer (5X) 8 L sol’n
   • 125 mM Tris-- 121 g
   • 960 mM Glycine-- 576 g
   • ddH2O
   • (SDS to be added individually to 0.1% of total sol’n)
10. Leupeptin
    • 5 mg container
    • 1000 ul ddH2O
    • dissolve and divide into 100 ul aliquots
    • freeze
11. MgCl2 (1 M) 1 L sol’n
    • 203 g MgCl2
    • ddH20
12. NaH2PO2-H2O (1 M) 500 ml sol’n
    • 69 g NaH2PO4-H20
    • ddH2O
13. Na3VO4 (200 mM) 100 ml sol’n
    • 3.6 g Na3VO4
    • ddH2O
14. Pepstatin (1 mg/ml)
    • 10 mg container
    • 10 ml methanol
    • dissolve and divide into 1 ml aliquots
    • freeze
15. PIPES (1M) 100 ml sol’n
    • 38 g PIPES
    • ddH2O
    • pH 6.8
16. PMSF
    • 1 g container
    • 30 ml isopropanol
    • dissolve (must be vortexed a great deal)
    • cover in alum. foil
    • store at room temp.
17. Ponceau S 200 ml sol’n
    • 1- 20 ml bottle Ponceau S Concentrated
    • 180 ml ddH2O
18. 2X Protein Sample Buffer
    • 2 M Tris, pH 6.8 --6.2 ml
    • 20% SDS --20 ml
    • Glycerol --20 ml
    • Bromophenol Blue --20 mg
    • ddH2O --54 ml
    • To Use: add 20% BME to volume of sample buffer needed (make fresh as needed)
19. Stripping Buffer (to strip and reprobe a filter) 50 ml sol’n
    • 62 mM Tris, pH 6.8 --1.5 ml
    • 2% SDS --5 ml 20% SDS
    • 0.7 ml BME/ 100 ml --350 ul
    • ddH2O
20. TAE (50 X) 1 L sol’n
    • 242 g Tris
    • 57.1 ml acetic acid
    • 100 ml 0.5 M EDTA (pH 8.0)
    • ddH2O
    • (dilute to 1 X before use)
21. TBS (10 X) 5 L sol’n
    • 500 mM Tris --302.5 g
    • 1.5 M NaCl --437.5 g
    • ddH2O
    • pH 8.0
22. TBS/ 5% BSA 20 ml sol’n
    • 2 ml 10X TBS
    • 18 ml ddH2O
    • 1 g BSA
23. TBS- Tween Sol’n
    • 500 ml 10 X TBS
    • 4.5 L ddH2O
    • 5 ml Tween 20
24. Transfer Buffer (for Protein Gels) 2 L sol’n
    • 28.5 g glycine
    • 6.1 g Tris
    • ddH2O