Sample dilution, Reagents, Gating – Luminex xPONENT 3.1 User Manual

Page 17: Plates

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5

Introduction

Sample Dilution

Dilute concentrated biological samples, such as plasma or serum, at least 1:5 with
reagents as part of assay setup or as a final dilution step. If running an xMAP-based kit,
follow the dilution instructions found on the kit’s product insert.

Reagents

Formulated reagents must be free of particulates other than xMAP beads. Do not dilute
xMAP calibrators or verifiers.

Gating

Gate positions are dependent upon buffer composition. Any changes made to the buffer
composition in an assay may result in a different optimal gate location.

Determine the gating on the Doublet Discriminator channel for the assay during assay
development. The numeric values appear on the left side of the histogram. Use the
numerical gate position, as determined during assay development, to set the gate location
in the protocol.

Gating information may change with a new lot of xMAP beads. Each time you receive a
new lot of xMAP beads, evaluate them with the current protocols. If gating information
changes, create a new protocol identical to the current protocol, but with a new version
number and new gating information. If running an xMAP-based kit, follow the instructions
found on the kit’s product insert or use the provided software protocol.

Plates

When using uncovered plates, use black opaque plates, if possible, to reduce
photobleaching.

For heated assays, use CoStar

®

Thermowell

®

96-well, thin-wall polycarbonate, model P

plates.

For nonheated assays, select a 96-well plate with an overall height no greater than 0.75
inches (19 mm). See “Bead Concentration” on page 3.

See the recommended consumables list on the Luminex website at http://
www.luminexcorp.com/support/recommendedmaterials/index.html
for more information.

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