Millipore - Antibody Recovery and Reuse With the SNAP i.d. Protein Detection System

Millipore Technical Publications

		AN- an1879en00
Antibody Recovery and Reuse With the SNAP i.d. Protein Detection System
Lit No:	an1879en00
Year:	2008

Many researchers reuse antibodies in western blotting. While many antibodies loose potency with time or even degrade due to improper storage conditions, it is important to recognize the potential value of recovering the primary antibody for possible re-use in some experiments. Here we show how the SNAP i.d. Protein Detection System can be used to collect antibodies for future reuse. Since this system is vacuum-driven, following the protocol outlined below is critical for successful antibody recovery.

Considerations for Successful Antibody Recovery

Antibody recovery is only possible in single and double well blot holder.
Position of the collection tray is critical for antibody recovery. For the single blot holder the tray should be positioned in the center of the manifold chamber. For the double well blot holder, the trays should be positioned with both trays’ ID tabs facing each other.
Before the addition of antibody, the manifold lid must be properly closed. Proper sealing is essential to ensure that the antibody will be collected in the trays.
In order to achieve maximum recovery, the system requires the addition of a small amount of buffer after the antibody incubation step. This is required to drive out antibody remaining in the blot holders. Although this step will result in a dilution of the collected antibody, it is essential for maximal recovery.

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