Q: I want to conjugate PJ25C activated allophycocyanin (APC) to an antibody. Do you have a recommended procedure?
A. We used to offer a conjugation kit (PJ25K) for small scale APC conjugations with up to 1mg Ab, that included PJ25C activated APC. PJ25K is now discontinued.
The details/recipes for all PJ25K conjugation kit buffers and spin columns are available in the PJ25K manual, together with a protocol.
We suggest following the PJ25K protocol, using PJ25C activated APC, with buffers you prepare and purification columns sourced using the specifications in the PJ25K manual. If this is successful, there is the option to scale up the procedure with more PJ25C APC.
Some monoclonal antibodies may precipitate upon DTT reduction used in the PJ25K protocol. An alternative protocol using SPDP/TCEP may be appropriate for antibodies that don’t fare well in DTT, and is provided in the following Tech Note:
We still carry an R-Phycoerythrin (R-PE )conjugation kit, PJ31K.
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