PhycoLink® R-Phycoerythrin Conjugation Kit
PhycoLink® R-Phycoerythrin (R-PE) Conjugation Kit contains everything you need to conjugate up to 1 mg (~1000 tests) of your antibody to RPE. Complete step-by-step protocols are included and allow small scale conjugations (down to 50 µg of antibody). This kit utilizes the best linking method and the brightest RPE available. Conjugates can be ready in as little as two hours.
Size: 1 kit
Product Code: PJ31K
Q. Can this conjugation kit be used to scale up to 10 mg of antibody?
Q. What happened to your KPK13 and KPK80 Conjugate Purification Kits?
A. KPK13 and KPK80 products were discontinued in October 2015. The KPK100 matrix used in KPK13 & KPK80 was Uniflow 6 (Sterogene) which separates based on size-exclusion chromatography (SEC). Uniflow 6 has been discontinued, but it has been replaced with Superflow 6 which has the same porosity and twice the flow rate. Superflow 6 or one of the other Sterogene SEC matrices should be a good alternative. Please contact Sterogene for details on the best SEC matrix for your application.
Another SEC option is Superdex 200 (GE Healthcare).
The purification matrix of choice may depend on the available equipment. The advantage of Uniflow/Superflow is that gravity-driven chromatography can be performed, whereas Superdex 200 requires a pump. If you have an FPLC or a suitable pump, Superdex is probably just as easy or maybe easier. If not, Uniflow/Superflow is the way to go without requiring new equipment.
Q. I need more of the desalting columns (A & B) that are supplied with your RPE conjugation kit (PJ31K). Where can I get them? Can they be re-used/regenerated? What about Spin Column D?
A. Desalting columns (Columns A & B) may be purchased from GE Healthcare (PD-10, catalog #17-0851-01). Initially we used columns from Pierce Chemical (catalog #43232), so you may see some difference in the appearance from each supplier. Column material is cross-linked dextran. Please note that the columns must be equilibrated before use.
Columns A & B may be re-used several times. To prepare a column for initial use or re-use, simply wash it with 5 - 10 column volumes of the appropriate buffer (recipes for buffers used in the Conjugation Protocol may be found in the Appendix of the PJ31K manual). We have used these columns 2 - 4 times over the course of a day, but we havent kept used columns for longer periods. You might be able to do so, but microbial growth is a concern.
Spin Column D may be purchased separately from Bio-Rad (Bio-Spin P-6 Gel Columns, catalog #7326002).
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: