Bio-Rad Bio-Gel P Polyacrylamide Gel User Manual

Page 7

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Fig. 5.1. Protein desalting with the Econo-Pac
10 DG desalting column.

9

5. Allow entire sample to enter the column. Add (3.0

ml - sample volume) of buffer. Allow buffer to enter
the column and discard the effluent.

6a. Add 1.5 x sample volume or 4ml of buffer

(whichever is less) to elute the higher molecular
weight component(s). Collect this fraction from the
column. For a more precise collection method refer
to step 6b.

6b. Elute the desired components with 8 ml of buffer,

while collecting 1 ml fractions from the column.
Analyze the fractions with the appropriate detector
and plot the results against the fraction number. Pool
the desired fractions (See Figure 5.1).

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Conditions
Column:

Econo-Pac 10 DG desalting column

Sample:

BSA in 250 mM NaCI

Buffer:

H

2

O

Flow rate:

1.0 ml/min

Peaks:
1. BSA
2. NaCI

Conductivity

O.D. 280

1

2

LIT81D 7/9/98 8:37 AM Page 8

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