Q: What species is this kit compatible with?
A: Our Rapid Antibody Purification Kit is a protein A column based purification kit that can be used to purify IgG from human, mouse, rabbit, pig, dog or cat.
Q: Can I use this kit with more than 5 mL of sample?
A: The capacity of the column is determined by the amount of IgG in the sample, which is listed on Table 1 of the product manual. It is fine to apply more than the recommended 5 mL, in successive additions of 5 mL at a time, as long as the maximum binding capacity is not exceeded. The column has a very high binding capacity, so it is unlikely that the column will be saturated.
Q: How can I improve the flow rate?
A: To get maximal binding with this kit, the antibody sample should be applied to the column by gravity flow. The flow rate of the column should be 1ml/min, which should be no longer than 10 minutes total for the wash step. If the wash is taking longer than this, there is likely an air bubble trapped inside the column. To remove any bubbles, cap the bottom of the column, add 2-3 mL of Binding Buffer to the top of the column, and replace the cap on the top of the column. Then either shake the column vigorously by hand for several seconds, or place the column in a 15 mL conical tube and centrifuge at 1000 g for 5 minutes.
Q: How should the purified antibody be stored?
A: The purified antibody can be stored at 4ºC; however long term storage conditions are antibody specific and should be determined for each antibody. This can be done by preparing aliquots and testing the antibody susceptibility to freeze/thaws at -20ºC or -80ºC.