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ANNOUNCE: NOSdetect assay kit from ALEXIS

Dr. Stephen Hoare s.hoare at alexisuk.demon.co.uk
Tue Apr 16 06:01:58 EST 1996

Cat. No. 850-006-KI01   1 Kit

ALEXIS now offers a nitric oxide synthase (NOS) assay kit, the NOSdetect
Assay Kit (Stratagene), to help researchers elucidate the many
biological functions of nitric oxide (NO), a molecule that is currently
the focus of intense investigations.

Bredt and coworkers [1] developed a simple, sensitive and specific assay
for NOS by monitoring the conversion of [3H]arginine to [3H]citrulline
as this conversion is stoichiometric with the enzymatic formation of NO
[2,3]. This approach has several advantages over other techniques and is
now the standard assay for NOS activity in crude and purified enzyme
preparations. First, the use of a radioactive substrate, [3H] or
[14C]arginine, makes the technique sensitive to the picomole range.
Second, the direct enzymatic conversion of arginine to citrulline in
eukaryotic cells is specific for the NOS pathway. Third, the facile
separation of citrulline from unreacted arginine permits hundreds of
assays to be performed in less than an hour.

[1] D.S. Bredt and S.H. Snyder; Ann.Rev.Biochem. 63. 175 (1994)
[2] D.S. Bredt and S.H. Snyder; PNAS 86, 9030 (1989)
[3] D.S. Bredt and S.H. Snyder; PNAS 87, 689 (1990)

Overview of the NOSdetect Assay Kit (Stratagene)
For routine assays, radioactive arginine is added to intact tissues or
protein extracts. After incubation, assays are stopped with a buffer
containing EGTA which chelates the required calcium, and applied to spin
columns containing cation-exchange resin to which arginine tightly
binds. Citrulline is ionically neutral at pH 5.5 and flows through the
column. The NOS activity is quantitated by counting the radio-activity
in the flowthrough.

- The assay is extremely sensitive (pM levels)

- The assay is rapid (results in 1 hour)

- The assay is easy to perform (simple to follow protocol)

- A detailed Technical Manual is included in each shipment

- The assay is useful for the measurement of iNOS and other isoforms

- The assay measures enzyme activity directly

Components and Storage Conditions
Cerebellum Extract (rat brain):  -80 C
Calmodulin Solution:  -20 C
Nw-Nitro-L-arginine methyl ester . HCl:  -20 C
2 x Reaction Buffer:  -20 C
All Other Components:  Room Temperature

For further information, please contact your local ALEXIS office.
Dr. Stephen Hoare, Alexis Corporation (UK) Ltd.
USA TEL: (619)658-0065      SWITZERLAND TEL: +41(0)62 299 29 20
=== FAX: (619)658-9224      =========== FAX: +41(0)62 299 24 80
 e-mail: alexis at primenet.com         e-mail: 101726.2030 at compuserve.com
UK  TEL: +44(0)1949 836111  GERMANY     TEL: +49(0)6401 900770
==  FAX: +44(0)1949 836222  =======     FAX: +49(0)6401 3823
 e-mail: alexis at alexisuk.demon.co.uk                       

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