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| Substances: | | ribonuclease; cytochrome c; lysozyme; lactoglobulin-b; ovalbumin |
| Column: | | NUCLEOSIL® 100-5C18 PPN, 125 x 4mm ID |
| Phase: | | Nucleosil 100-5 C18 PPN |
| MN catalog number: | | |
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| Sample pretreatment: | | alkaline eluent through the column as shown below. |
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| Conditions: | | eluent A: 0.1% TFA in H2O, eluent B: 0.08% TFA in CH3CN, gradient from 20 to 60% B in 10min
flow rate 1.0ml/min |
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Detection
Subsequent analysis: | | UV, 280nm |
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| Author(-s): | | MACHEREY-NAGEL |
| Source: | | LC department |
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| Keywords: | | proteins |
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Chromatogram:
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| Legend: | | Separation of standard proteins
1. ribonuclease
2. cytochrome c
3. lysozyme
4. b-lactoglobulin
5. ovalbumin
Left: before treatment, right: after treatment with 300 column volumes of the alkaline eluent at a flow rate of
0.4ml/min (alkaline eluent = 1 volume 50mM NaOH + 1 volume n-propanol, saturated with SiO2 by stirring the mixture with 1g/l unmodified silica gel for 1day). |