Avidin biotin amplified immunoperoxidase staining of angiotensin‐1‐converting enzyme transferred to nitrocellulose after agarose isoelectric focusing

Abstract
Angiotensin‐1‐converting enzyme (ACE) subjected to isoelectric focusing in agarose gel can be transferred to nitrocellulose in a reliable and reproducible manner. Specific detection of ACE is enhanced 2‐fold by replacing enzyme‐conjugated reagents with avidin biotin‐conjugated reagents. Sensitivity is not diminished when the carcinogenic substrate 3,3′‐diaminobenzidine is replaced with the non‐carcinogenic substrate 3‐amino‐9‐ethylcarbazole. This improved procedure was used to monitor the spontaneous change in the isoelectric point of ACE which occurred during purification of ACE from outdated human plasma.

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