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Multiplex Immunoassays

The Animal Phenotyping Platform uses the Luminex xMAP Intelliflex DR-SE analyzer for high-throughput immune-based protein assays.
Luminex multiplex analysis

This allows multiplex measurements to be performed, enabling several analytes to be determined simultaneously in the same sample volume.

The analytes are quantitatively fluorescently labeled and the emitted fluorescence is measured.

The required sample volume can be very small, often less than 5 µl, maximum 50µl.

For example, a standard assay can be run on a 96-well plate and is designed to analyze 80 serum or plasma samples. The multiplex analyzer used at the APP, Luminex xMAP Intelliflex DR-SE, offers the ability to measure in 96-well plates as well as in the large 384-well approach.

Commercially available assays, specific for mouse and rat, among others, are available for a variety of analytes and address research fields e.g. in immunology, cancer, cardiovascular, metabolism, development and many more.

 

Quantitatively fluorescently labeled analytes

 

Method

 

In the multiplex approach, the targeted proteins are immunologically coupled to color-labeled beads. The use of classification lasers (bead lasers) and reporter lasers enables the simultaneous identification of proteins and their respective quantification. The laser optics ensure highest precision and data density of the measurements.

Simulation of the identifaction of the labeled proteins
Assay examples

 

Several established assays from different vendors are available for mouse and rat, e.g.:

Mouse Cytokine/Chemokine Panel (32-plex):

Eotaxin/CCL11 | IL-1β | G-CSF | GM-CSF | IFNγ | IL-1α | IL-1β | IL-2 | IL-3 | IL-4 | IL-5 | IL-6 | IL-7 | IL-9 | IL-10 | IL-12 (p40) | IL-12 (p70) | IL-13 | IL-15 | IL-17A/CTLA8 | IP-10/CXCL10 | KC/GRO/CINC-1/CXCL1 | LIF | LIX | MCP-1/CCL2 | M-CSF | MIG/CXCL9 | MIP-1α/CCL3 | MIP-1β/CCL4 | MIP-2/CXCL2 | RANTES/CCL5 | TNFα | VEGF-A

Mouse Cardiovascular Panel (9-plex):

sCD40L | CXCL16 | Endocan-1 | Follistatin | LIGHT | Oncostatin M | Placental Growth Factor 2 | TnI | TnT

Mouse Adipokine (7-plex)

IL-6 | Insulin | Leptin | MCP-1/CCL2 | PAI-1 | Resistin | TNFα

Mouse/Rat Pituitary Gland (7-plex):

ACTH | BDNF | FSH | GH | LH | Prolactin | TSH

Rat Stress Hormone (3-plex):

ACTH | Corticosterone | Melatonin

Rat Bone Panel (8-Plex):

ACTH | DKK1 | FGF-23 | Insulin | Leptin | Osteoprotegerin | PTH | Sclerostin