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Page Title:
SETTING UP THE LUMINOMETER (Dynex MLX 1000 Luminometer) |
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 ERDC TN-DOER-C10
March 2000
Substrate B, PHARMINGEN #556870 - 1 aliquot (reconstituted).
Cell Lysis Buffer, PHARMINGEN #556871 - 1 aliquot (1x) .
50-mL solution basins, nonsterile.
Vortex (with microplate adaptor holder).
Inverted microscope.
Countertop Centrifuge (with microplate adaptor holder).
Luminometer (Dynex MLX 1000; DYNEX Technologies, Chantilly, VA).
96-well luminometer plate, opaque, nonsterile (Microlite 2.0, Dynex # 7417).
B.
REAGENT PREPARATION.
The reagents for the P450RGS assay can be prepared ahead of time and stored in
aliquots in the -80 C freezer. Contents and date of preparation should be properly
labeled and initialed on these reagent containers.
1) Substrate A.
a) Reconstitute lyophilized Substrate A with 50 mL of ultrapure water.
b) Transfer 12.5-mL aliquot of Substrate A solution into each 15-mL tube.
c) Store tubes in -80 C freezer.
2) Substrate B (Light Sensitive).
a) Reconstitute lyophilized Substrate B with 50 mL of ultrapure water.
b) Transfer 12.5-mL aliquot of Substrate B solution into each 15-mL tube.
c) Wrap each tube with aluminum foil. Substrate B is light sensitive.
d) Store tubes in -80 C freezer.
3) Cell Lysis Buffer (1x).
a) Pipet 100 mL of ultrapure water into 50 mL of the 3x cell lysis buffer in a
beaker.
b) Transfer 12.5 mL of the 1x cell lysis buffer in 15-mL tubes.
c) Store tubes in -80 C freezer.
C.
SETTING UP THE LUMINOMETER (Dynex MLX 1000 Luminometer).
1) Turning on and initializing the luminometer.
a) Turn on the power switches of the luminometer (bottom-right corner of the
luminometer) and monitor. The luminometer will run a self test. The results
of the test will be displayed on the self-test window. The Revelation
window will then be displayed.
b) Click on the Utility menu, then the Dispensers option, followed by the
Initialize option.
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