RNA was extracted from 500 [micro]L plasma and the
elutant was 50 [micro]L.
1 M H3PO4 as mobile phase and the
elutant was scanned at 208 nm; for IAA, 35% methanol prepared in 0.
The material submerged in 1 mL of PBS and 1 mL and
elutant were collected and replaced every 10 min for the first hour and then hourly for the next 8 hours.
The
elutant was dried using anhydrous sodium sulphate; once dry, the supernatant was transferred to a conical-base graduated tube, a small amount of dry [N.
Eighty percent cold acetone fractionation of pooled
elutant fractions of Agarose column chromatography concentrates the protein, acetone precipitation selected as priority over ammonium sulphate saturation since the latter imparts brownish colour to extract even after dialysis against 0.
Electrophoresis: Samples of
elutant from the Dynabead affnity method were prepared for gel electrophpresis by mixing 1:1 v/v in Laemmli was performed on 4-15% gels (BioRad) in TRIS/ glycine/SDS buffer (Biorad cat# 161-0732) for one hour st 120 V.
We also collected the
elutant or flow-through sample containing nonadherent cells from the sP tubes.