A Range of Composite Sorbents for Size Exclusion of Biological Macromolecules
- High resolution
- Semi-rigid particles
- Sorbents feature narrow particle size distribution and narrow pore size distribution
- Four different types are available, allowing fractionation of molecules with molecular weights ranging from 1,000 to 350,000 daltons
- Suitable for small-, medium-, and large-scale applications
- Fractionation and purification of biological molecules by size exclusion
- Determination of molecular weights
- Desalting of proteins and nucleotides
|
| Particle Size (µm) |
60 - 140 |
60 - 140 |
60 - 140 |
60 - 140 |
| Acrylamide (%) |
3 |
4 |
5 |
20 |
| Agarose (%) |
4 |
4 |
4 |
2 |
| Exclusion Limit (Da) |
750,000 |
200,000 |
90,000 |
22,000 |
| Linear Fractionation Range (Da) |
20,000 - 350,000 |
10,000 - 130,000 |
5,000 - 70,000 |
1,000 - 15,000 |
| Resolution Power (plates/m) |
1,500 |
1,500 |
1,500 |
3,000 |
| Desalting Capacity |
- |
- |
- |
45% gel vol. |
| Working pH |
3 - 10 |
3 - 10 |
3 - 10 |
3 - 10 |
| Storage Temperature |
2 - 8 ° C (36 - 46 ° F) |
|
Determination of a Selectivity Curve for Ultrogel AcA 54 Sorbent Using a Mixture of Known Molecular Weight Proteins
Column dimensions: 1.6 x 40 cm; Buffer: 0.05 M Tris-HCl, pH 7.4 containing 0.17 M sodium chloride: sample constituted of blue-dextran 2000 (a) for the determination of the void volume(Vo); bovine serum albumin (b) (MW 68,000); ß-lactoglobulin (c) (MW 35,000); Myoglobin (d) (MW 17,800); Cytochrome c (e) (MW 12,400); Sample volume: 0.6 mL; Flow rate: 4.8 cm/h. The insert represents the elution profile of the above protein standards from the above column.
Large-Scale Desalting on Ultrogel AcA 202 Sorbent
Column dimensions: 5 x 37 cm; volume: 730 mL; Sample: bovine serum albumin (5 mg/mL) containing NaCl (6.5 mg/mL); Flow rate: 7 cm/h (i.e. 140 mL/h). Sample volumes: A = 36 mL (5% of sorbent volume); B = 220 mL (30% of sorbent volume); C = 327 mL (45% of sorbent volume); 280 nm UV detection.
Gravity Flow Column Desalting With Ultrogel® AcA 202 Sorbent
Desalting in a gravity drip column format was evaluated with Ultrogel AcA 202 sorbent against competitive agarose and dextran-based pre-packed columns following manufacturers’ directions. The resulting desalting curves plotting protein (absorbance at 280 nm) and salt elution (m or μS/cm, measured using a Horiba micro-volume conductivity meter) are summarized below. The results all show excellent separation between the excluded protein peak and the retained salt. Ultrogel AcA 202 gave the best separation between the void fraction containing the proteins of interest and the salt or included peak elution, suggesting that a higher volume of sample could have been loaded on this 10 mL column compared to other sorbents.
Gravity flow (10 mL) columns were poured with Ultrogel AcA 202 sorbent and two competitive agarose and dextranbased media. A 1 mL protein sample [5 mg/mL human serum albumin (HSA) in 1 M NaCl] was then applied and fractions of 1 mL collected after elution with high purity water. Absorbance was measured at 280 nm and conductivity was measured using a Horiba micro-volume meter.
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