Table 5.
Standards for hydrodynamic analysis
| Protein | M r aa seq | S 20,w | S max /S | R s (nm) | Source | M r S-M |
|---|---|---|---|---|---|---|
| Ribonuclease A beef pancreas | 14,044 | 2.0a | 1.05a | 1.64 | HBC | 13,791 |
| Chymotrypsinogen A beef pancreas | 25,665 | 2.6 | 1.21 | 2.09 | HBC | 22,849 |
| Ovalbumin hen egg | 42,910s | 3.5 | 1.27 | 3.05 | HBC | 44,888 |
| Albumin beef serum | 69,322 | 4.6a | 1.33 | 3.55 | S-M, HBC | 68,667 |
| Aldolase rabbit muscle | 157,368 | 7.3 | 1.45 | 4.81 | HBC | 147,650 |
| Catalase beef liver | 239,656 | 11.3 | 1.21 | 5.2 | S-M | 247,085 |
| Apo-ferritin horse spleen | 489,324 | 17.6 | 1.28 | 6.1 | HBC | 451,449 |
| Thyroglobulin bovine | 606,444 | 19 | 1.37 | 8.5 | HBC | 679,107 |
| Fibrinogen, human | 387,344 | 7.9 | 2.44 | 10.7 | S-M | 355,449 |
Gel filtration calibration kits, containing globular proteins of known molecular weight and Rs, are commercially available (GE Healthcare, Sigma-Aldrich). These same proteins can be used for sedimentation standards. The proteins in these kits are included in the table along with some others that we have found useful. The values for Mr given in the first column are from amino acid sequence data. Values for S20,w and Rs are from the Siegel–Monte paper (indicated S-M under source), or the CRC Handbook of Biochemistry [3] (indicated HBC). They agree with the values listed for Rs in the GE Healthcare gel filtration calibration kit, with the exception of ferritin. The "Mr calc" in the last column was obtained by our simplification of the Siegel–Monte calculation (M = 4,205 S Rs). Note that the worst disagreement with "Mr aa seq" is about 10%
aS for ribonuclease A is questionable because of the low Smax/S (1.05). S values for bovine serum albumin vary in the literature from 4.3 to 4.9. Many sources use 4.3, but we find that 4.6 gives a better fit with other standards (note that the standard curve in Figure 5 used 4.3, but 4.6 would have placed it closer to the line)