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MiniExcellose is resin column for purification of much protein (>10mg) such as GST, MBP, His tagged fusion protein by gravity. The procedure is brief, allowing completion of the protocol in 1 hour.
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1. Remove the suspension of column, and add 6ml of equilibration buffer to the column.
2. Remove the end cap and flow the equilibration buffer through the column.
3. Repeat 1,2.
4. Load the protein sample into the column, and collect the flow through fractions.
5. Wash the column with 6ml of equilibration buffer, and collect the wash fractions.
6. Repeat 5.
7. Elute with 3ml elution buffer, and collect the eluted fractions.
8. Collect the each fractions (4,5,7) and these aliquots will be electrophoresed.
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Comment
1. The kinds and amounts of buffer described in this manual are commended.
2. If necessary, adjust the buffer conditions according to user¡¯s purpose.
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Column Storage
1. After experiment, wash the column with equilibration buffer.
2. After washing with 20% ethanol, add 4ml of 20% ethanol, and cover with column cap.
3. Remove the end cap, close the column cap and then lid the end cap to prevent forming air bubbles in the column.
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| Catalog No |
Product |
Total binding Capacity |
Equilibration buffer |
Elution buffer |
Vol.
(ml) |
24101
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IDA MiniExcellose ¢ç
|
120uM Cu 2 + |
50mM phosphate, 0.5N NaCl, pH 8.0 |
Equilibration buffer +
0.5M imidazole |
3 |
| 24201 |
GST MiniExcellose ¢ç |
10mg GST |
PBS, pH 7.4 |
Equilibration buffer + 10mM reduced glutathione |
3 |
24301
|
MBP MiniExcellose ¢ç |
20mg MBP |
20mM Tris, 200mM NaCl, 1mM EDTA 10mM 2-mercaptoethanol, pH 7.4 |
Equilibration buffer + 10mM maltose |
3 | |
