Binding buffer: 20 mM sodium phosphate, 150 mM NaCl, pH 7.4
Elution buffer I: 100 mM sodium citrate, pH 6.0
Elution buffer II: 100 mM sodium glycine, pH 2.7
Neutralization buffer: 1M Tris-HCl, pH 9.0
Buffers mentioned above are needed to be prepared by customers!
1) Wash the prepacked 5 ml column with 5~10 column volumes of distilled water to remove 20% ethanol.
2) Equilibrate the column with 5~10 column volumes of binding buffer.
3) Filtrate 3~5 ml target sample through a 0.45 m filter and load the sample.
4) Wash with 5~10 column volumes of binding buffer.
5) Wash with 10 column volumes of elution buffer I and followed by 10 column volumes of elution buffer II.
6) Discard first 1/3 column volume of elution buffer II and immediately collect 1 column volume of eluant by elution buffer II and neutralize collect fractions with neutralization buffer.
7) Analyze the elute fraction to ensure that target molecule is captured (Figure 1).