During my ProcartaPlex assay data analysis, I am getting a warning message that there is high bead aggregation. What should I do?
Here are possible causes and solutions for this issue:
- Check the protocol settings (make sure you select the correct DD settings).
- Check the level of sheath fluid and empty the waste.
- Before acquiring the plate, run calibration and verification beads on the Luminex instrument.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
During my ProcartaPlex assay data analysis, the beads fall below or to the lower left of their bead region on the bead map. Why is this?
Here are possible causes and solutions for this issue:
This usually indicates that the beads have been photobleached. This problem can also be caused by exposing the beads to organic solvents. Unfortunately, the assay will have to be repeated because the beads cannot be restored. The beads must be protected from light and organic solvents.
Alternatively, the instrument may be off in its measurements or you may have a calibration issue. Call the manufacturer for a service appointment.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
During my ProcartaPlex assay data analysis, beads do not appear in the region gated. What happened?
This indicates that an incorrect buffer was used for the final step. The Wash Solution provided in the kit must be used for washing the beads and the Reading Buffer should be used for resuspending the beads before loading them into the Luminex instrument. The osmolarity of the solution will impact the size of the bead, and any change in the bead size will alter detection by the instrument.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
The bead counts for all of my ProcartaPlex assay wells are erratic. What went wrong?
Here are some suggestions:
- Before acquiring the plate, run calibration and verification beads on the Luminex instrument.
- Review the instrument settings and make sure they are appropriate for the assay being run (adjustment of needle height, make sure you select the correct bead gates and the correct DD settings).
- Shake the plate before acquisition on the instrument to resuspend the beads.
- Vortex the beads for 30 sec before adding them into the plate.
- Washing: Do not forget to keep the plate for about 2 mins on the Hand-Held Magnetic Plate Washer before emptying the plate.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
The bead count for my ProcartaPlex assay standard curve is correct and regular. The bead count for my samples is erratic and I get a warning message saying "The acquisition had at least one region that did not reach the maximum count". What should I do?
This pattern is indicative of a sample matrix effect. Here are some suggestions:
- Confirm that the sample has been clarified and is free of debris and free of lipids (5-10 min centrifugation recommended).
- Confirm that there is at least a 1:1 ratio of sample to assay diluent for serum, plasma samples. For cell lysates or tissue homogenates, confirm that the sample has been diluted appropriately in assay buffer to reduce the concentration of detergent in the lysis buffer to ⋜0.01%. For other sample types, further sample optimization may be required.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
