Bio-Rad Affi-Gel 15 Gel User Manual
Page 7
Table 1. Protein Coupling to Affi-Gel 10 and
Affi-Gel 15 Support
Coupling Efficiency (%)
Protein
pl
Affi-Gel 15 Gel
Affi-Gel 10 Gel
1. Fetuin
3.3
43
3.0
2. Alpha-1-antitrypsin
4.0
76
5.0
3. Ovalbumin
4.7
70
8.5
4. Bovine serum albumin
4.9
80
14
5. Human transferrin
5.9
87
36
6. Bovine hemoglobin
6.8
59
83
7. Human globulin
5.8-7.3
39
90
8. Myoglobin
6.8-7.8
10
85
9. Cytochrome c
9.3
0
90
10. Lysozyme
10-11
1
95
9
Fig. 2. Protein coupling with Affi-Gel and Affi-Gel 15 sup-
ports.
Coupling conditions: Each protein solution (3 ml 0.1 M
MOPS, pH 7.5, containing 40 mg protein) was combined with
2 ml of Affi-Gel media. The gel slurry was mixed at 4 °C for 2
hours, and then stripped with 7 M urea containing 1 M NaCl.
The uncoupled protein was determined, using published
E
1 cm
, by dilution of an aliquot of the urea effluent into 0.1 M
HCl and measurement of the absorbance at 280 nm (
●—●
Affi-Gel 15 gel;
●
●
—
●
●
Affi-Gel 10 gel).
0
10
20
30
40
50
60
70
80
90
100
% Protein Coupled
0
5
10
15
20
mg coupled
ml gel
3
4
5
6
7
8
9
10
11
pl
8
280
LIT156B 6/17/98 10:41 AM Page 8