• UV-visible spectrophotometer and disposable cuvettes
• Mini-Protean 3 vertical gel electrophoresis rigs
• Power supplies
• Hot plates

• Biological Macromolecules
  • Lactalbumin, bovine milk, MW 14,175 Da
  • Lysozyme, chicken egg white, MW 14,307 Da
  • Myoglobin, horse heart, MW 16,900 Da
  • Chymotrypsin, bovine pancreas, MW 21,600 Da
  • Trypsin, bovine pancreas, MW 23,281 Da
  • Aldolase, rabbit muscle, MW 39,188 Da per subunit
  • Alpha Amylase, Bacillus species, MW ~50,000 Da

• Tris free base
• HCl
• Bovine Serum Albumin (BSA) Standard, 2 mg/mL
• Bradford Reagent
• Solutions for Electrophoresis
  • 30% Acrylamide/Bisacrylamide Solution (CAUTION: Neurotoxin)
  • 1.5M Tris-HCl buffer, pH 8.8
  • 0.5M Tris-HCl buffer, pH 6.8
  • 20% (w/v) Sodium Dodecyl Sulfate (SDS)
  • 10% (w/v) Ammonium Persulfate (APS)
  • TEMED
  • 4X Tris-Glycine Electrophoresis Buffer (dilute to 1X before use)

• Solutions for Coomassie Staining and Destaining
  • Methanol
  • Acetic Acid
  • Coomassie Brilliant Blue R-250
• Gel Drying Solution (prepared by the TA)\

Abbreviations:

• mcL = microliters
• dI = deionized

1.5M Tris-HCl buffer at pH 8.8

• Calculate the amount in grams of Tris free base (MW = 121.14 g/mol) required for 50 mL of 1.5M Tris base solution. Verify the accuracy of your calculation with the TA. Dissolve the appropriate amount of Tris base in <50 mL of dI water.
• Measure the pH of the Tris base solution. Adjust the pH to 8.8 using HCl. (CAUTION: HCl is a strong acid. Wear gloves when handling this reagent and do not inhale this reagent.)
• Do not exceed a total solution volume of 50 mL!
• Add additional dI water as necessary to adjust the final solution volume to 50 mL.
• Mix the solution. Label a storage container with the reagent name, concentration, measured pH, your name and the date of preparation. This reagent may be kept stored at room temperature.

0.5M Tris-HCl buffer at pH 6.8

• Calculate the amount in grams of Tris free base (MW = 121.14 g/mol) required for 25 mL of 0.5M Tris base solution. Verify the accuracy of your calculation with the TA. Dissolve the appropriate amount of Tris base in <25 mL of dI water.
• Measure the pH of the Tris base solution. Adjust the pH to 6.8 using HCl. (CAUTION: HCl is a strong acid. Wear gloves when handling this reagent and do not inhale this reagent.).
• Do not exceed a total solution volume of 25 mL!
• Add additional dI water as necessary to adjust the final solution volume to 25 mL.
• Mix the solution. Label a storage container with the reagent name, concentration, measured pH, your name and the date of preparation. This reagent may be kept stored at room temperature.

20% (w/v) Sodium Dodecyl Sulfate (SDS)

• Dissolve 1g of SDS in dI water to a final volume of 5 mL.
• This reagent may be stored at room temperature. Crystals may form after long-term storage. Resolubilize any crystallized SDS by heating prior to use.

10% (w/v) Ammonium Persulfate (Prepare fresh)

• Weigh out 0.1g of ammonium persulfate directly into a 1.5 mL microcentrifuge tube in an analytical balance.
• Add 1 mL of dI water using a P1000 pipettor.

Preparation of 4X Electrophoresis Buffer

• Dissolve 12g of Tris free base, 57.6g of glycine, and 4g of SDS in dI water to a total volume of 1 L.
• At the time of use, dilute this solution 4-fold (to 1X) by adding 250 mL of 4X electrophoresis buffer to 750 mL of dI water.

Preparation of Staining and Destaining Solutions

• Combine 125 mL of methanol, 25 mL of glacial acetic acid, and 100 mL of dI water to make 250 mL of destaining solution.
• Dissolve 0.1g of Coomassie Brilliant Blue R-250 in 50 mL of the destain solution to make 50 mL of Coomassie staining solution.