New amiloride analogue as hapten to raise anti-amiloride antibodies.
A new amiloride analogue, "amiloride-caproic acid," was synthesized, coupled to albumin, and used as a hapten to raise anti-amiloride antibodies in rabbits. The antibodies were affinity purified with an amiloride affinity column and characterized. Binding studies using [3H]benzamil showed a dissociation constant of 0.8 nM. Amiloride and amiloride-caproate inhibited [3H]benzamil binding; epsilon-guanidinocaproic acid showed no inhibition. Anti-amiloride antibodies reversed the inhibition by amiloride of sodium transport across toad urinary bladder. Anti-amiloride antibodies and an amiloride affinity column should provide useful tools for the characterization of the epithelial sodium channel.
Kleyman
T R
TR
Rajagopalan
R
R
Cragoe
E J
EJ
Erlanger
B F
BF
Al-Awqati
Q
Q
D000418:Albumins; D000584:Amiloride / Q000031:analogs & derivatives* / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D001667:Binding, Competitive; D001693:Biological Transport, Active / Q000187:drug effects; D002024:Bufo marinus; D005260:Female; D006146:Guanidines / Q000378:metabolism; D006241:Haptens; D005779:Immunodiffusion; D011817:Rabbits; D012964:Sodium / Q000378:metabolism; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism
D000418:Albumins; D000906:Antibodies; D006146:Guanidines; D006241:Haptens; C013407:benzamil; C047657:amiloride caproate; C047656:epsilon-guanidinocaproate; D000584:Amiloride; D012964:Sodium