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Fig. 1. Separation of standard.

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.

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

0 10 20 30

Minutes

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