Commonly Used Buffers
Protein | Subunit molecular weight | pI | Protein | Subunit molecular weight | pI |
Myosin, heavy chain (rabbit muscle) | 205 | 6,45, 6,7 | Carbonic anhydrase (bovine erythrocytes) | 30,0 | 5,85 |
â-Galactosidase | 116 | 4,4 | Trypsinogen, PMSF-treated | 24,0 | 9,3 |
Phosphorylase b (rabbit muscle) | 97,4 | 6,3 | Trypsin inhibitor (soybean) | 20,0 | 4,55 |
Bovine serum albumin | 67 | 4,9 | â-Lactoglobulin | 18,4 | 5,2 |
Catalase | 57,5 | 6,7 | Myoglobin (sperm whale) | 16,8 | 6,8, 7,2 |
Ovalbumin | 5 | 4,7, 4,9 | Lysozyme (hen egg white) | 14,3 | 11 |
Aldolase | 40 | 8,9 | á-Lactalbumin | 14,2 | 4,4 |
Pepsin | 34,7 | 2,8 to 4,03 |
Molecular Weights and pl Values of Common Protein Standards
Buffer | Stock solution | Components | Amount | Conc. in to use* solution | Final conc. stock in 1X solution |
PBS (Phosphate Buffered Saline) | 10X | NaCl | 80g | 1,37M | 137mM |
KCl | 2g | 27mM | 2,7mM | ||
Na2HPO4 7H2O (sodium phosphate dibasic) | 11,5g | 43mM | 4,3mM | ||
KH2PO4 (potassium phosphate monobasic) | 2g | 14mM | 1,4mM pH is 7,3 |
||
SSC (Saline–Sodium Citrate) | 20X | NaCl | 175g | 3M | 150mM |
Na3C6H5O7 2H2O (sodium citrate) | 88g Adjust pH of stock to 7,0 Adjust pH to 7,5 |
0,3M | 15mM | ||
TAE (Tris–Acetate–EDTA) | 50X | Tris base | 242g | 2M (Tris Acetate) | 40mM |
Acetic acid, glacial | 57,1mL | ||||
0,5M EDTA, pH 8,0 | 37,2g | 0,1M | 2mM | ||
pH of stock is 8,5 | |||||
TBE | 10X | Tris base | 108g | 0,89M | 89mM |
(Tris–Borate– | Boric acid | 55g | 0,89M | 89mM | |
EDTA) | 0,5M EDTA, pH 8,0 | 40mL | 20mM | 2mM | |
TE (Tris–EDTA) | 1X | Tris base | 1,2g | 10mM | 10mM |
0,5M EDTA, pH 8,0 | 2mL | 1mM Adjust pH to 7,4, 7,5, or 8,0 |
1mM |
* Amounts given are to make 1L of stock solution.
Approximate pH Values for Various Concentrations of Stock Solutions
Reagent | 1N | 0,1N | 0,01N | 0,001N |
Acetic acid | 2,40 | 2,90 | 3,40 | 3,90 |
Hydrochloric acid | 0,10 | 1,07 | 2,02 | 3,01 |
Sulfuric acid | 0,30 | 1,20 | 2,10 | |
Citric acid | 2,10 | 2,60 | ||
Ammonium hydroxide | 11,80 | 11,30 | 10,80 | 10,30 |
Sodium hydroxide | 14,05 | 13,07 | 12,12 | 11,13 |
Sodium bicarbonate | 8,40 | |||
Sodium carbonate | 11,50 | 11,00 |
SDS-PAGE Stock Solutions
Stock Solutions | Catalogue No | Amount to use | Special instructions |
30% Acrylamide/0.8% Bis-Acrylamide | |||
Acrylamide1 | BPE170 | 30,0g | Filter stock solution through a 0,45ìm to 100mL filter and store at 4°C in the dark. Discard after thirty days as acrylamide gradually hydrolyzes to acrylic acid and ammonia |
Bis-Acrylamide | BPE171 | 0,8g | |
H2O | — | ||
4x Tris•Cl, pH 6.8 (0.5M Tris•Cl) | |||
Tris Base | BPE152 | 6,05g | Dissolve Tris Base in 40mL H2O, then adjust pH to 6,8 with 1N HCl. Add H2O to 100mL total volume. Filter through a 0,45ìm filter store at 4°C. |
H2O | — | 40mL | |
1N HCl | — | as needed | |
H2O | — | to 100mL | |
4x Tris•Cl, pH 8.8 (1.5M Tris•Cl) | |||
Tris Base | BPE152 | 91g | Dissolve Tris Base in 300mL H2O, then adjust pH to 8,8 with 1N HCl. Add H2O to 500mL total volume. Filter through a 0,45ìm filter store at 4°C. |
H2O | — | 300mL | |
1N HCl | — | as needed | |
H2O | — | to 500mL | |
10% SDS | |||
SDS | BPE166 | 50g | — |
H2O | — | to 500mL |
1- Caution: Acrylamide monomer is neurotoxic. Wear gloves while handling, and do not pipet by mouth.