14 MagPix Protocol
14.1 Overview
This protocol will support you to set up a run on the xPONENT software, as was performed for this study. This protocol is used to test specimens with the final 12-plex assay on the MagPix machine, as well as to carry out the confirmations of bead conjugation and the cross-reactivity tests,
Please note that this protocol does not detail basic operations, routine maintenance and calibration of the MagPix. Please refer to the manufacturer’s guidance on this.
14.2 Setup
- In the xPONENT software, go to ‘Protocols’ > ‘Create new Protocol’.
- Enter a name for the protocol e.g. “12-plex Biomarker” and select the appropriate plate type from the drop-down menu.
- Set the sample volume to 50 µL. Click ‘Next’.
- Select and label the required bead regions with their coupled antibodies so that the table on screen maps to the table below:
| Name | Analysis | Units | Count | Region |
|---|---|---|---|---|
| sTREM1 | No Analysis | 50 | 14 | |
| Ang2 | No Analysis | 50 | 15 | |
| Ang1 | No Analysis | 50 | 18 | |
| sTNFR1 | No Analysis | 50 | 20 | |
| IL6 | No Analysis | 50 | 21 | |
| IL8 | No Analysis | 50 | 22 | |
| TRAIL | No Analysis | 50 | 25 | |
| IL10 | No Analysis | 50 | 26 | |
| Azu | No Analysis | 50 | 27 | |
| MxA | No Analysis | 50 | 28 | |
| IP10 | No Analysis | 50 | 29 | |
| CHI3L1 | No Analysis | 50 | 30 |
- Click ‘Next’
14.3 Plate Layout
Highlight all wells and select ‘Unknown’ so that the whole plate is yellow.
Ensure that ‘Direction’ is the left-hand option for vertical reading order.
Add a ‘Post-Batch Routine’ under ‘Commands and Routines’.
Click ‘Save’.
The protocol is ready to use and can now be found under ‘Protocols’.
We have used this uniform plate layout for all plates, but it is also possible to add standards with their concentrations, and blanks, to the layout if this is desirable.
We recommend uploading or adding sample (or well) IDs to each plate (‘batch’) before running a plate so that raw data contains sample IDs.
NOTE - Adding standards to the plate layout may cause the output csv file to have additional rows and produce errors if used with our R analysis script. The analysis script should be modified in the relevant places to accommodate this change if you wish to add standards.