Jhdavis::Immunoprecip: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(New page: Immunoprecipitation (sherman et al. 1992 EMBO J 11: 71-77) 20 mL culture, mid-log phase centrifuge 5 min, 5000 rmp resuspend in 2 mL of 10 mM tris pH 8.0, 1 mM EDTA, 1mg/mL lysozyme freez...) |
(No difference)
|
Revision as of 13:29, 30 April 2009
Immunoprecipitation (sherman et al. 1992 EMBO J 11: 71-77)
20 mL culture, mid-log phase centrifuge 5 min, 5000 rmp resuspend in 2 mL of 10 mM tris pH 8.0, 1 mM EDTA, 1mg/mL lysozyme freeze/thaw sonication centrifuge 10 min, 14,000 rmp dilute 50 uL sup with 950 uL PBS add antibody and incubate 30 min, 4 C add proteinA-sepharose (35 uL of 1:1 beads:PBS suspension) 1.5 h at 4 C centrifuge 1 min, 1000 rpm wash 3 times with PBS, 1% tritonx100, 5 mM EDTA
