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I would like to use this Serum Purification kit on an antibody that is 60 uL of Ascites fluid. Should I dilute this antibody to a volume of 5mL? Will the purified antibody be at an acceptable concentration for use with the HRP conjugation kit or will I need to concentrate it further?
Asked on Mar 28 2012
Thank you for your phone calls.
I have heard back from the lab and they recommend using the purification kit ab102784 for small volumes of tissue culture supernatant or ascites fluid (100-500 uL). When using this kit, it will be fine to dilute your 60 uL of ascites fluid to 100 uL with PBS.
The purified antibodies can then be used with our EasyLink conjugation kits. For optimal results, the concentration of the antibody should be between 0.5-5 mg/mL. You may still be able to conjugate the antibody at lower concentrations, but it will likely not be as efficient and may result in unlabeled antibody present in the solution.
If you switch to the purification kits for smaller volumes, the concentration will likely not be a problem, but if you dilute your antibodies for use with the TCS or Ascites purification kits, you may need to concentrate it before use with the EasyLink kits. This can be done by using our concentration spin columns ab102776 or ab102778.
Since you have already purchased the TCS and Ascites purification kits, I will be happy to arrange an exchange if you would like to switch to the kits designed for smaller volumes. Let me know if you need any other information or assistance.
Answered on Mar 28 2012