DESCRIPTION Recombinant ribonuclease
inhibitor effectively inhibits ribonuclease A type (ribonuclease A, B, and
C) enzyme activity by binding non-covalently to RNase molecules at a 1:1
ratio. Recombinant ribonuclease inhibitor has the same inhibitory activity
as the natural protein from human placenta but devoid of human DNA or
proteins.
SOURCE
Recombinant strain of E. coli.
APPLICATIONS ¡Ü Protection of mRNA in
cDNA synthesis. ¡Ü Improvement of protein translation performance.
¡Ü Increased yields and activity of polysomes. ¡Ü Improvement of in
vitro virus replication. ¡Ü Improvement of RNA translation in
homologous systems.
UNIT DEFINITION One unit is defined as
the amount of ribonuclease inhibitor required to inhibit by 50% the
activity of 5ng of ribonuclease A.
STANDARD UNIT ASSAY CONDITIONS Activity
is measured by the inhibition of hydrolysis of cytidine 2',3'-cyclic
monophosphate by ribonuclease A, where 200 units of ribonuclease inhibitor
produces 50% inhibition of 1¥ìg of ribonuclease A.
CONCENTRATION: 20-40 units/¥ìl.
STORAGE CONDITIONS 20mM HEPES-KOH
(pH7.6), 50mM KCl, 8mM DTT, and 50% glycerol. Store at -20¡É for frequent
use and -70¡É for long term storage. If stored for long period of time or
opened frequently, addition of DTT to 5mM final concentration into the
reaction mixture is recommended to maintain optimal activity.
NOTE Recombinant ribonuclease inhibitor
is active over a broad pH range but requires a minimum of 1mM DTT to
maintain activity.
REFERENCES 1. Blackburn, P. et al.
(1977) J. Biol. Chem. 252, 5904. 2. Blackburn, P. and Jailkhani, B. L.
(1979) J. Biol. Chem. 254, 12488.
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