Beamline phone numbers:
+44 (0) 1235 77 8627
+44 (0) 1235 77 8640
Principal Beamline Scientist:
Nathan Cowieson
Tel: +44 (0) 1235 567 504
E-mail: [email protected]
Email: [email protected]
Tel: +44 (0)1235 56 7675
SEC-SAXS experiments are the gold standard experiment for high quality monodisperse data. A typical protein SEC-SAXS sample is 50 µL at 1-10 mg/ml, with 500 mL of buffer usually shipped a 10x stock.
Column |
Dimensions (mm) |
Volume (mL) |
Size Range (Da) |
pH range |
Pressure limit (Bar) |
Flow rate (mL/Min) |
Run time (Mins) |
SRT-C SEC-150 (Sepax) |
4.6 x 300 |
4.99 |
500-150,000 |
2-8.5 |
170 |
0.16 |
32 |
SRT-C SEC-300 (Sepax) |
7.8 x 300 |
14.34 |
5,000-1,250,000 |
2-8.5 |
170 |
0.3 |
50 |
SRT-C SEC-300 (Sepax) Replacing KW-403 (Shodex) |
4.6 x 300 |
4.99 |
5,000-1,250,000 |
2-8.5 |
170 |
0.16 |
32 |
SRT-C SEC-500 (Sepax) |
4.6 x 300 |
4.99 |
15,000-5,000,000 |
2-8.5 |
130 |
0.16 |
32 |
KW-402.5 (Shodex) |
4.6 x 300 |
4.6 |
5,000-100,000 |
3-7.5 |
80 |
0.16 |
32 |
KW-404 (Shodex) |
4.6 x 300 |
4.6 |
30,000-4,000,000 |
3-7.5 |
80 |
0.16 |
32 |
KW-405 (Shodex) |
4.6 x 300 |
4.6 |
100,000-20,000,000 |
3-7.5 |
80 |
0.16 |
32 |
Superdex 30 (Cytiva) |
3.2 x 300 |
2.4 |
100-7000 |
3-12.0 |
30 |
0.075 |
32 |
Superdex 75 (Cytiva) |
3.2 x 300 |
2.4 |
3,000-70,000 |
3-12.0 |
30 |
0.075 |
32 |
Superdex 200 (Cytiva) |
3.2 x 300 |
2.4 |
10,000-600,000 |
3-12.0 |
30 |
0.075 |
32 |
Superose 6 (Cytiva) |
3.2 x 300 |
2.4 |
20,000-5,000,000 |
3-12.0 |
30 |
0.075 |
32 |
Buffers will be optimised to stabilise the sample. Including 1-2% glycerol or sucrose helps reduce radation damage. Including some salt (i.e. 100-150 mM) will help reduce interation between the particle and the column and inter-particle correlation effects that could reduce data quality. Detergents/lipids can form higher order structures that can interact with the sample and are best avoided.
There are some things to consider when planning your SEC-experiment:
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