Performance of multistep immunochemical reactions by counterflow isotachophoresis on nitrocellulose membranes--I. Immunoblotting

Molecular Immunology
G I Abelev, E R Karamova

Abstract

Urine with trace amounts of different proteins from healthy people or B-lymphoma patients was concentrated and separated simultaneously by counterflow isotachophoresis on cellulose acetate membranes (CAM). The protein zones were blotted onto nitrocellulose membrane (NCM) by direct contact of CAM and NCM. NCM-blots were exposed to second isotachophoresis with the leading electrolyte 0.06 M Tris-HCl and the terminating one, 0.012 M Tris-beta-alanine. Under these conditions the moving boundary formed by Cl-/beta-alanine- migrated towards the anode with decreasing velocity. At a certain point the rate of migration of the moving boundary became completely compensated by the electroendosmotic counterflow. In this steady state position the boundary stopped on the NCM support, while the electroendosmotic rate in the area before the boundary was much higher than the rate of the opposite migration of any protein to the anode. Under these conditions electroendosmosis served as a "conveyer belt" which transferred consecutively the immunoreagents, antibodies, immunoconjugates, or antiperoxidase-peroxidase system through the protein blots "printed" on NCM. The immunoblots obtained in this way were developed by the substrate for the immunoenz...Continue Reading

References

May 1, 1987·Biulleten' eksperimental'noĭ biologii i meditsiny·A I FaermanA Iu Koliada
Aug 15, 1986·International Journal of Cancer. Journal International Du Cancer·M J EscribanoP Burtin
Sep 4, 1984·Journal of Immunological Methods·H Towbin, J Gordon
Nov 1, 1984·Analytical Biochemistry·G I Abelev, E R Karamova

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Citations

Jun 1, 1991·Electrophoresis·D SchranzG I Abelev
Sep 27, 2014·Lab on a Chip·Tally Rosenfeld, Moran Bercovici
Dec 20, 2018·Lab on a Chip·Tally Rosenfeld, Moran Bercovici
Aug 25, 2011·Analytical Chemistry·Jos QuistHeiko J van der Linden

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