Bio-Rad Profinia™ Protein Purification Instrument - Manual (Page 130)

Table 10.4. Formulations for buffers and solutions provided in the GST kits.
Profinia Supplied Concentrated Working 1x Vol, Buffer
Solution Formulation Formulation ml Position
GST lysis 2x 300 mM NaCl, 150 mM NaCl, 125 N/A
buffer 20 mM Na
2
HP0
4
, 10 mM Na
2
HP0
4
,
10 mM EDTA,pH 7.4 5 mM EDTA, pH 7.4
GST 2x 300 mM NaCl, 150 mM NaCl, 200 1
wash 20 mM Na
2
HP0
4
, 10 mM Na
2
HP0
4
,
buffer 10 mM EDTA, pH 7.4 5 mM EDTA, pH 7.4
GST 2x 40 mM glutathione, 20 mM glutathione, 100 3
elution 200 mM Tris, 10 mM EDTA, 100 mM Tris, 5 mM EDTA,
buffer pH 8.0 pH 8.0
Desalting 5x 685 mM NaCl, 13.5 mM KCl, 137 mM NaCl, 2.7 mM KCl, 200 4
buffer 21.5 mM Na
2
HP0
4
, 4.3 mM Na
2
HP0
4
,
40.5 mM KH
2
P0
4
, pH 8.1 mM KH
2
P0
4
, pH 7.4
7.0 (pH 7.4 upon dilution)
Cleaning 2x 1,000 mM NaCl, 100 mM Tris, 500 mM NaCl, 50 mM Tris, 125 5
solution 1 pH 8.0 pH 8.0
Cleaning 4x 2,000 mM NaCl, 500 mM NaCl, 125 6
solution 2 400 mM NaOAc, pH 4.5 100 mM NaOAc, pH 4.5
Storage 2x 4% C
6
H
5
CH
2
OH (benzyl 2% C
6
H
5
CH
2
OH (benzyl 200 7
Solution alcohol) alcohol)
Figure 10.3 shows proper placement of GST buffers in the Profinia instrument.
Fig. 10.3. GST buffers properly positioned with instrument lid closed.
10.5 Desalting Buffer and Separation Kits
The buffers and solutions provided in the desalting kit consist of a PBS-based desalting
buffer, cleaning buffers, and a storage solution. The PBS solution provides an optimum
ionic strength to prevent nonspecific interactions between proteins and the matrix, and is a
commonly used buffer for the storage of a wide variety of proteins.
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