# User:Catherine Koenigsknecht/Notebook/Experimental Biological Chemistry/2011/09/21

Biomaterials Design Lab <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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## Entry title

Continue to purify using the column.

## Objective

Learn how to maintain an OpenWetWare Notebook.

## Description

1. Run 50mL of the supernatent from yesterday over the column at 1mL/min. Collect 10mL fractions.
2. Wash the column with 70mL of column buffer at 1mL/min. Collect 10mL fractions.
3. Elute the column with 25mL of 10mM maltose in column buffer at 1mL/min. Collect 5mL fractions.
4. Run 150mL of the supernatent from yesterday over the column at 1mL/min. Collect 10mL fractions.
5. Wash with 75mL of column buffer at 1mL/min. Collect 10mL fractions.
6. Elute the column with 75mL of 10mM maltose in column buffer at 1mL/min. Collect 5mL fractions.

## Data

• Add data and results here...

## Notes

• 10mM maltose(100mL) = 0.36g maltose + column buffer
• We did both of our washes until the "Bradford-by-eye" revealed that no more protein was being washed off of the column.
• A "Bradford-by-eye" is mixing 20μL of Bradford dye with 60μL of water and 20μL of a fraction. When the mixture is bright blue protein is present, a lighter blue means there is less protein, and when the mixture is orange little to no protein is present.

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