A new technique for detecting oxytocinase activity in electrophoresis gels.

A new technique for detecting oxytocinase activity in electrophoresis gels.

Electrophoresis (1992-06-01)

A C Roy, N Saha, S M Tan, F Z Kamarul, S S Ratnam

ABSTRACT

A sensitive staining method for the detection of oxytocinase (EC 3.4.11.3) activity in electrophoresis gels has been described. The method is based on the enzymatic release of p-nitroaniline (PNA) from two specific synthetic oxytocinase substrates, S-benzyl-L-cysteine-p-nitroanilide (BCN) and L-leucine-p-nitroanilide (LN), respectively. The PNA is then diazotized with sodium nitrite and subsequently coupled to a chromogen, N-(1-naphthyl)-ethylenediamine dihydrochloride (NED) to produce a deep pink/magenta colored azo-dye at the site of oxytocinase activity.

MATERIALS

Product Number

Brand

Product Description

N9125

Sigma-Aldrich

N-(1-Naphthyl)ethylenediamine dihydrochloride, ≥98%

70720

Supelco

N-(1-Naphthyl)ethylenediamine dihydrochloride monomethanolate, for spectrophotometric det. of nitrate and nitrite, ≥99.0%

33461

Supelco

N-(1-Naphthyl)ethylenediamine dihydrochloride, for determination of sulfonamide and nitrite, ACS reagent, ≥98%

222488

Sigma-Aldrich

N-(1-Naphthyl)ethylenediamine dihydrochloride, ACS reagent, >98%