I will be performing ELISA on patient blood that was discovered to be stored at 4C for about 5 weeks. I was unable to attain standard separation upon centrifugation at 2500rpm for 10min. What settings, if any, would be useful to separate antibodies at this stage?
Wolfgang Schechinger
You could directly dilute the samples with PBS or similar for ELISA. If cells and cell debris (membranes, nuclei, …) are a problem, pass the samples through a centrifugal filter. If you want to extract total IgG first, use a Protein A/G column.
If lipids are causing problems, use petrol ether to remove them.
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