Protein G beads for IP - (Oct/03/2007 )
Hey there I am a little new to the lab scene and could use someones judgement.
I have 100mg of dry Protien G beads that I hydrated in 1XM PBS +. I was told to store the hydrated beads in a one to one ratio with 1XPBS +. My hydrated beads were approximately 500ul so I put 500 ul of PBS and stored at 4C.
I am running an IP using 600, 000 cells, and approx 5ul of primary antibody. My question is about how much protein G solution (G with PBS) should I add to have a good pull down.
Thanks a ton
Do you know the concentration of your antibody?
Manual of protein G beads tells you the IgG binding capacity!
It's just a simple calculation!