Implen P 300 User Manual

NanoPhotometer
®
P-Class User Manual
Version 2.1 Page 22 / 70
4.2.3 Protein UV Dye Method
The procedure is as follows:
Parameter Screen
NanoVolume Applications
Cuvette Applications
Parameter Screen
Step 1 Press 1 for NanoVolume OR 2 for Cuvette folder.
Step 2 Press 2 to select Protein folder.
Step 3 Press 2 to select Protein dye mode.
Step 4 Using NanoVolume Applications select the Lid Factor as
described in the “Average Detection Range Sheet” and
under 3.2. A minimum of 1.5 µl sample volume (for lid
10) is recommended.
Step 5 Enter the Dilution Factor using the keypad numbers.
Range 1.00 to 9,999. Use the C button to backspace and
clear the last digit entered. OR press Menu/Options to
enter the dilution factor screen. Enter the volume of the
sample using the keypad numbers. Range 0.01 to 9,999.
Enter the volume of the diluent using the keypad
numbers. Range 0.01 to 9,999. Press OK to calculate
the dilution factor and return to the Parameters screen
OR press Cancel to cancel the selections and return to
the Parameters screen.
Step 6 Select whether the Dye correction (calculation of the dye-
dependent correction factor) is used or not with the left
and right arrows. The Background correction is always
calculated in the Dye methods.
Step 7 Select the Protein (BSA (default), Serum Albumin
(mouse), Serum Albumin (human), IgG, Lysozyme,
Custom or OD 1).
Step 8 If using Custom Protein there are two possibilities to
enter the correct factors (see also page 18 protein UV
method):
Molar extinction coefficient (M
-1
* cm
-1
):
Ranges are:
Wavelength: 200 nm to 340 nm
Molar extinction coefficient (M
-1
* cm
-1
): 10,000 to
9,999,999
Molecular weight: 0.001 to 9,999,999
Extinction coefficient (l/g * cm):
Ranges are:
Wavelength: 200 nm to 340 nm
Extinction coefficient (l/g * cm): 0.001 to 9,999
Step 9 Select the Units of measurement using the left and right
arrows. Options: mg/ml, μg/ml, ng/μl and μg/μl.
Step 10 Enter the protein-dependent extinction coefficient. Range
is 10,000 to 9,999,999.
Step 11 Press OK to store the chosen parameters and to enter
the next screen OR Cancel
to return to the Protein
folder.
Step 12 Select the appropriate Dye Type. 4 different AlexaFluors,
2 Cy-Dyes, 2 DyLight Dyes, FITC, Pacific Blue, r-PE and
Texas Red are programmed with their corresponding
maximum absorbance wavelength, dye-dependent
extinction co-efficient and dye-dependent correction
factor at 280 nm. For further details please refer to 12.4
Protein fluorescent dye incorporation.
Step 13 If using Custom Dye maximum absorbance wavelength of
the custom dye, dye-dependent extinction coefficient and
dye-dependent correction factor at 280 nm have to be
entered. Ranges are:
Dye Abs Max: 300 nm to 950 nm
Dye Ext. Coefficient: 10,000 to 9,999,999
Dye Correction: 0 to 0.999

Implen P 300 Specifications

General

Brand	Implen
Model	P 300
Category	Measuring Instruments
Language	English