00:53Preparation of NE Depleted of U1 snRNP (U1ΔNE)
02:10Immunoprecipitation of 10S Fractions of Glycerol Gradients by Anti-Gal3
03:52Assembly of Splicing Reaction and Analysis of Products
06:09Results: RNA and Polypeptide Compositions of the Anti-Gal3 Precipitate of the 10S Region of the Glycerol Gradient and its Ability to Reconstitute Splicing in U1ΔNE
This article describes the experimental procedures for (a) depletion of U1 snRNP from nuclear extracts, with concomitant loss of splicing activity; and (b) reconstitution of splicing activity in the U1-depleted extract by galectin-3 - U1 snRNP particles bound to beads covalently coupled with anti-galectin-3 antibodies.