Beckman Coulter Biomek FX-ADMETox Skrócona instrukcja obsługi - Strona 2

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4. Prepare test compounds
.
a.Spectroscopic purity DMSO for test compound
preparation.
b.Add DMSO to test compounds according to the table
below:
Test Compounds
MW
Weight (mg)
DMSO (ml)
Ketoprofen
254.3
2.54
Verapamil
491.1
4.91
Antipyrine
188.2
9.40
Metoprolol
684.8
6.85
Carbamazepine
236.3
2.36
Ranitidine
350.9
3.51
Propanolol
295.8
2.96
c.Add 80 µl of each test compound solution into each
well of the StockPlate.
DMSO
Ketoprofen
Verapamil
Antipyrine
Metoprolol
Carbamazepine
Rantidine
Propranolol
d.Extra test compound can be frozen for future use.
5. Populate the worksurface with appropriate labwares.
a.Place 3 96-well UV plates on worksurface position.
b.Place Biomek P200 tip boxes on worksurface position.
c.Place PAMPA Sandwich Plate (pre-loaded) on
worksurface position.
d.Place SUPPORT Plate on worksurface position.
e.Place the StockPlate on worksurface position.
f.Place the LipidPlate on worksurface position.
g.Place ASB solution Low Profile Reservoir on
worksurface position.
h.Place System Buffer High Profile 12 Trough Reservoir
on worksurface position.
i.Place Deep well plate for diluted test compounds on
worksurface position.
6. Enter test compound information into Excel template file.
a.Launch Microsoft Excel application.
b.Select File | Open.
c.Browse to open PAMPA Template.xlt file located in
the PAMPA Evolution command Software directory.
d.Enter each compound in the same well location that it
was loaded in the StockPlate.
e.Physiochemical constants are optional.
f.Do not leave blank spaces in the Excel template sheet.
Concentration (mM)
g.Rename the Excel file and save in DATA sub-folder in
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the PAMPA Evolution command Software.
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h.Exit the Excel application.
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7. Run PAMPA assay.
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a.Click the Import from Excel button on the toolbar.
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b.The Input Compounds dialog box appears, click the
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Browse button to import test compound Excel
template file.
c.Click the OK button for the Warning dialog box for
correct StockPlate type.
d.Verify the loaded compound names and
physiochemical constants, click the OK button to
continue.
e.Open the Instrument-Settings, select the Use pre-
measured blank.
f.Browse to select DefaultBlankPlate.spc.
g.Uncheck Tell me about low-UV wells...
h.Click the OK button and exit the Instrument Settings.
i.Click the General Information button, fill in the
appropriate experimental conditions, click the OK
button.
j.Click the Start Assay button.
k.Select the Uninterrupted Assay Flow and Use pre-
measured blank check boxes.
l.Verify that Double-Sink Protocol is selected, and
incubation time is set to "0.5 hr".
m.Select Properties and verify the System Solution
Volume = 600 µL and Stirring be ABL = 40 µM.
n.Select Multi pH Assay and select pH Map.
o.Verify the updated layout of System Solution at
different pH.
p.Click the OK button and Start the assay to proceed.