How to Perform Glycan Analysis and Generate GU Values in Empower - Tip225

OBJECTIVE or GOAL

Tip #225: How to Perform Glycan Analysis and Generate GU Values in Empower

In this week’s tip, we will learn how to perform Glycan analysis and generate GU values that can be used in the screening of Glycans. This example demonstrates the analysis with Glycans that has been labeled with a fluorescent tag. (Please Note: You must have the GPC option for your Empower system to use this procedure.)

ENVIRONMENT

• Empower

PROCEDURE

STEP 1
In the sample set, select ‘Inject Narrow Standards’ for the Dextran Ladder Standard and ‘Inject Broad Unknows’ for the Samples (figure 1).

STEP 2
With the row for the ‘Dextran Ladder Standard’ highlighted, click the ‘Amount’ tool, select ‘Molecular Weights’ tab for the current sample and enter the values for each peak in the Dextran Ladder. (Please Note: The number of values entered must match the number of integrated peaks in the chromatogram) (figure 2).

STEP 3
Create a GPC processing method, setting up peak detection and integration parameters for the narrow standard and the broad samples (figure 3).

STEP 4
On the Calibration tab, Select Relative for the GPC technique, 5th order fit type, time for the x-axis and low to high for the calibration order (figure 4).

STEP 5
Create a simple peak custom field which divides the peak molecular weight by 10000 and call it GU or GU Value (figure 5).

STEP 6
Process the Sample Set and view the result for the Dextran Ladder standard (figure 6).

STEP 7
View the resulting calibration curve (figure 7).

STEP 8
View the results for a sample (figure 8).

 

It’s that easy!

Final Note: You can use either the Pro or QuickStart interface for this tip.
 

ADDITIONAL INFORMATION