Bio-Rad Chromatography Standards - Manual (Page 3)

Fig. 1. Separation of standard.
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Protein Standard
Components
Component Molecular Weight
Cytochrome c 12,000
Myoglobin (horse) 17,000
Lysozyme 14,000
Recommended Volume
of Standard
Figure 1 shows separations of the standard using Hi-Pore ®
RP-304 HPLC columns. A volume of 20 µl standard was
applied to each column. The protein peaks give absorbance at
280 nm from about 0.03 to 0.1 absorption units.
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Conditions
Column:
Reversed Phase
Hi-Pore RP-304, 250 x 4.6 cm
Sample: 20 µl
Eluant: A. 0.1% TFA
B. 0.1% TFA, 95% acetonitrile
25-50% B linear gradient, 30 min
Flow Rate: 1.5 ml/min
Detection: UV Monitor @ 280 nm
0.08 AUFS
Peaks:
1. Cytochrome c
2. Myoglobin
3. Lysoz
yme
2
3
1
0 10 20 30
Minutes
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LIT162B 10/28/98 9:20 AM Page 2