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Not for use in diagnostic or therapeutic procedures. Not for resale.
Problem
Variation between
duplicate samples.
Low bead number
in samples.
High background.
Little or no detection
of protein in sample.
Less than six bead
populations are observed
during analysis or
distribution is unequal.
Debris (FSC/SSC) during
sample acquisition. Also
for plasma samples.
Overlap of bead population
fluorescence (FL3)
during acquisition.
Standards assay tubes
show low fluorescence
or poor standard curve.
All samples are positive
or above the high standard
mean fluorescence value.
Vortex Capture Beads before pipetting. Beads can aggregate.
Avoid aspiration of beads during wash step. Do not wash or resuspend beads
in volumes higher than recommended volumes.
Test various sample dilutions, the sample may be too concentrated.
Remove excess Mouse Inflammation PE Detection Reagent by increasing the
number of wash steps as the background may be due to non-specific binding.
Sample may be too dilute. Try various sample dilutions.
Ensure that equal volumes of beads were added to each assay tube.
Vortex Capture Bead vials before taking aliquots. Once Capture Beads
are mixed, vortex to ensure that the beads are distributed evenly throughout
the solution.
Increase FSC threshold or further dilute samples.
Increase number of wash steps if necessary. Make a tighter FSC/SSC region
gate around the bead population.
This may occur in samples with very high protein concentration.
Ensure that instrument settings have been optimized using the Cytometer
Setup Beads.
Check that all components are properly prepared and stored.
Use a new vial of standard with each experiment and once reconstituted,
do not use after 12 hours.
Ensure that incubation times were of proper length.
Dilute the samples further. The samples may be too concentrated.
Suggested Solution
Biohazardous samples.
It is possible to treat samples briefly with 1% paraformaldehyde before
analyzing on the flow cytometer. However, this may affect assay performance
and should be validated by the user.
analyzing on a flow cytometer.
model. Direct quantitation of proteins from the rat model has
not been validated using this kit and results may vary.
