Bio-Rad Profinia™ Protein Purification Instrument - Manual (Page 42)

The Bio-Rad methods function provides two sample loading flow rate options:
• Standard — recommended for normal- to high-expression proteins (<100 kD)
• Low — may improve the yield of low-expression proteins, high MW proteins (>100 kD)
or samples containing detergents
The Bio-Rad methods function provides two cartridge wash options:
• Standard — recommended for most purification conditions
• Extended — may improve the purity of product for some sample types
Parameters for each type of method are provided in Table 6.1.
Table 6.1. Sample loading Flow rates and wash volumes by cartridge size for each method
type option.
Method Type Cartridge Sample Loading Wash-1
Size, ml Flow Rate, ml/min Column
Volume (CV)
Standard Low Standard Extended
IMAC, Native 1 2 0.5 6 10
5 10 2.5 6 10
IMAC, Denaturing 1 2 0.5 6 10
5 10 2.5 6 10
GST, 1 1 0.5 12 15
Protein A and G
5 5 2.5 12 15
eXact 1 1 – 14 –
5 5 – 10 –
To select the flow rate(s) and wash time(s) for your sample(s):
1. Touch the arrow button to select either the “Standard” or “Low” flow rate for each
sample.
2. Touch the arrow button to select either the “Standard” or “Extended” wash option for
each sample.
Note: If a method will be performed with two samples, make flow rate and wash time
selections separately for each sample.
When the method options are selected, touch the Next button to proceed to the Enter Run
& Sample Information screen. A View Method button will be available in the lower toolbar to
view the individual method steps for the sample(s).
6.3.2 Selecting Sample Incubation Time
Each eXact Bio-Rad method is available with an option to select from three different incubation
times, 0.5 hrs, 2.0 hrs, or 15 hrs (Figure 6.7).
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