Comparison of oriented and random antibody immobilization in immunoaffinity chromatography of cytokinins.
Immunosorbents for the plant hormones cytokinins prepared by random antibody immobilization (to Affi-Gel 10) and by oriented approach via oxidized carbohydrate moieties on the Fc region (to Affi-Gel Hz or hydrazide derivative of Perloza MT 200) have been compared. Both approaches yielded immunosorbents with high dynamic capacity (ca. 5-10 nmol ml gel-1). Oriented antibody immobilization did not exhibit crucial effects in the case of low-molecular-mass cytokinins. Antibodies immobilized via a spacer to Affi-Gel 10 have probably enough conformational freedom to enable good accessibility to cytokinins. The sorbents were used in analysis of endogenous cytokinins in maize seeds. In phosphatase treated samples trans-zeatin and its riboside were predominant.
Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
R
R
Vankova
A
A
Gaudinová
H
H
Süssenbeková
P
P
Dobrev
M
M
Strnad
J
J
Holík
J
J
Lenfeld
D000906:Antibodies / Q000737:chemistry*; D002482:Cellulose; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003583:Cytokinins / Q000737:chemistry*; D007120:Immunochemistry; D007074:Immunoglobulin G / Q000737:chemistry; D008968:Molecular Conformation; D010084:Oxidation-Reduction; D012639:Seeds / Q000737:chemistry
D000906:Antibodies; D003583:Cytokinins; D007074:Immunoglobulin G; D002482:Cellulose