I need to do a pull-down/ IP with a purified zz tagged protein. In a reference paper they used IgG sepharose (brand not mentioned). I would like to go for magnetic beads, I saw you can use dynabeads M-270 epoxy and conjugate IgG. Anyone know other options from different companies and/or ready made beads?
Hello
We used a ZZ tagged protein and purified it with IgG coupled 1.0 µ microspheres by PolyLink (using the coupling kit:
http://www.polysciences.com/Catalog/Department/Product/98/categoryid--84/productid--1782/search--24350-1/).
IgG-Sepharose or -Agarose coupled resin is not working properly due to its porosity.
If you don't mind the price, Invitrogen might sell certain secondary antibody-coated dynabeads (used to sell under Dynal). Make sure you have the right species, some animal antibodies don't have strong affinity to protein A (or ZZ domain in your case). Some best ones are human and rabbit.
Thanks guys. On the mean time I found this protocol from Rockefeler U that I decided to test: http://prowl.rockefeller.edu/protocols/conjugation_magnetic_beads.html
I have a related question, can I use just Rabbit IgG as a primary antibody to probe for zz containing proteins in E. coli lysates by western blot or is it too dirty?
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