Chromatin Immunoprecipitation: Difference between revisions

Iharvey (talk | contribs)
Iharvey (talk | contribs)
No edit summary
Line 110: Line 110:
supernatant fraction.
supernatant fraction.
# Wash the Protein G Agarose-antibody/chromatin complex by resuspending the beads in 1 mL each of the cold buffers in the order listed below and incubating for 3-5 minutes on a rotating platform followed by brief centrifugation (3000-5000 x g for 1 minute) and careful removal of the supernatant fraction:
# Wash the Protein G Agarose-antibody/chromatin complex by resuspending the beads in 1 mL each of the cold buffers in the order listed below and incubating for 3-5 minutes on a rotating platform followed by brief centrifugation (3000-5000 x g for 1 minute) and careful removal of the supernatant fraction:
a. Low Salt Immune Complex Wash Buffer (Catalog # 20-154), one wash
* Low Salt Immune Complex Wash Buffer (Catalog # 20-154), one wash
b. High Salt Immune Complex Wash Buffer (Catalog # 20-155), one wash
* High Salt Immune Complex Wash Buffer (Catalog # 20-155), one wash
c. LiCl Immune Complex Wash Buffer (Catalog # 20-156), one wash
* LiCl Immune Complex Wash Buffer (Catalog # 20-156), one wash
d. TE Buffer (Catalog # 20-157), two washes
* TE Buffer (Catalog # 20-157), two washes
D. Elution of Protein/DNA Complexes
D. Elution of Protein/DNA Complexes
Prior to starting this section:
Prior to starting this section: