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BAYLOR NIAID Luminex Core: Service
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General Information
Each 96-well microplate can hold one 8-point standard curve and 40 samples run in duplicate. If you have two different media types or sample types that require a second standard curve on the same plate, then there is room for a total of 32 samples divided in the way that meets your needs. All analytes are purchased by the core only after requested due to variable pre-plex panels and custom panels built just for you.
Sample Preparation
For cell culture supernatants, please preclear your samples. There should be no cells present in the supernatant. The samples may be provided in either microcentrifuge tubes or 96-well microplate format. Please avoid multiple freeze-thaw cycles. We will also need 15ml of your cell culture media for each sample type in order to dilute samples and standard curve dilutions as per protocol. Serum samples are prepared and provided in microcentrifuge tubes or cryovials.
Volumes
We need a minimum of 50ul of your cell culture supernatant or serum sample. This volume allows for duplicate wells. If volume required cannot be provided, an increased dilution factor may be utilized but then the low-end sensitivity may be lost if there are only low levels of protein.
Human Luminex Panels
(Updated April 7, 2006)
| Bio-Rad |
Chemicon |
R&D |
Invitrogen |
| |
IL-1a |
IL-1a (Panel A) |
IL-1a |
| IL-1ra |
IL-1ra |
IL-1ra (Panel A) |
IL-1ra |
| IL-1b |
IL-1b |
IL-1b (Panel A) |
IL-1b |
| IL-2 |
IL-2 |
IL-2 (Panel A) |
IL-2 |
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sIL-2Ra |
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IL-2R |
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IL-3 |
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IL-3 |
| IL-4 |
IL-4 |
IL-4 (Panel A) |
IL-4 |
| IL-5 |
IL-5 |
IL-5 (Panel A) |
IL-5 |
| IL-6 |
IL-6 |
IL-6 (Panel A) |
IL-6 |
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sIL-6R |
| IL-7 |
IL-7 |
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IL-7 |
| IL-8 |
IL-8 |
IL-8 (Panel A) |
IL-8 |
| IL-9 |
IL-9 |
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| IL-10 |
IL-10 |
IL-10 (Panel A) |
IL-10 |
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IL-12p40 |
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IL-12p40 |
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IL-12p40/p70 |
| IL-12p70 |
IL-12p70 |
IL-12p70 (Panel B) |
IL-12p70 |
| IL-13 |
IL-13 |
IL-13 (Panel B) |
IL-13 |
| IL-15 |
IL-15 |
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IL-15 |
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IL-16 |
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IL-16 |
| IL-17 |
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IL-17 (Panel A) |
IL-17 |
| |
IL-18 |
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| G-CSF |
G-CSF |
G-CSF (Panel A) |
G-CSF |
| GM-CSF |
GM-CSF |
GM-CSF (Panel A) |
GM-CSF |
| IFNg |
IFNg |
IFNg (Panel A) |
IFNg |
| TNFa |
TNFa |
TNFa (Panel A) |
TNFa |
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TNFb |
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Eotaxin |
Eotaxin (Panel B) |
Eotaxin |
| MCP-1 |
MCP-1 |
MCP-1 (Panel A) |
MCP-1 |
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MCP-2 |
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MCP-3 |
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MCP-3 |
| MIP-1a |
MIP-1a |
MIP-1a (Panel A) |
MIP-1a |
| MIP-1b |
MIP-1b |
MIP-1b (Panel A) |
MIP-1b |
| Rantes |
Rantes |
Rantes (Panel A) |
Rantes |
| IP10 |
IP10 |
IP10 (Panel B) |
IP10 |
| VEGF |
VEGF |
VEGF (Panel A) |
VEGF |
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ENA-78 (Panel A) |
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| FGF-2(basic) |
FGF-2(basic) |
FGF-2(basic)
(Panel A) |
FGF-2(basic) |
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EGF |
EGF (Panel B) |
EGF |
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HGF (Panel B) |
HGF |
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IFNa |
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IFNa |
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MIG |
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Flt-3 Ligand |
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PDGF-AA |
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C-reactive Protein |
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PDGF-AB/BB |
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| PDGF-BB |
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PDGF-BB |
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BDNF |
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sFas Ligand |
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GDNF |
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Aß40 |
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Aß42 |
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GROa |
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GROa |
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MDC (CCL22) |
(MMP-1, -2, -3, -7, -8, -9, -12, -13 available) |
(MMP-3, -9, -13 3Plex) |
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TNF-RI |
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TNF-RII |
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sICAM-1 |
Thrombopoietin
(Panel A) |
DR5 |
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sVCAM-1 |
CD40 Ligand
(Panel B) |
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CXCL11/I-TAC
(Panel B) |
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Leptin (Panel B) |
Leptin |
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TGF-ß1,2,3 (cannot be plexed with any other analyte) |
** Panel A and B cannot be plexed together** |
Adiponectin |
Mouse Luminex Panels
(Updated April 7, 2006)
| Bio-Rad |
Chemicon |
R&D |
Invitrogen |
| IL-1a |
IL-1a |
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IL-1a |
| IL-2 |
IL-1b |
IL-1b |
IL-1b |
| IL-2 |
IL-2 |
IL-2 |
IL-2 |
| IL-3 |
IL-3 |
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| IL-4 |
IL-4 |
IL-4 |
IL-4 |
| IL-5 |
IL-5 |
IL-5 |
IL-5 |
| IL-6 |
IL-6 |
IL-6 |
IL-6 |
| KC |
KC |
KC |
KC |
| IL-9 |
IL-9 |
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| IL-10 |
IL-10 |
IL-10 |
IL-10 |
| IL-12p40 |
IL-12p40 |
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IL-12p40/p70 |
| IL-12p70 |
IL-12p70 |
IL-12p70 |
IL-12p70 |
| IL-13 |
IL-13 |
IL-13 |
IL-13 |
| IL-17 |
IL-17 |
IL-17 |
IL-17 |
| GM-CSF |
GM-CSF |
GM-CSF |
GM-CSF |
| IFNg |
IFNg |
IFNg |
IFNg |
| TNFa |
TNFa |
TNFa |
TNFa |
| MCP-1 |
MCP-1 |
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MCP-1 |
| Rantes |
Rantes |
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VEGF |
VEGF |
VEGF |
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MIP-2 |
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IP10 |
| Eotaxin |
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| G-CSF |
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G-CSF |
| MIP-1a |
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MIP-1a |
| MIP-1b |
MIP-1b |
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MIP-1b |
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MIP-3b |
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JE/CCL2 |
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Leptin |
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Resistin |
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Adiponectin/Acrp30 |
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MIG |
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FGF-2 (Basic) |
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PDGF-BB |
Rat Luminex Panels
(Updated April 7, 2006)
| Bio-Rad |
Chemicon |
R&D |
Invitrogen |
| IL-1a |
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IL-1a |
| IL-1b |
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IL-1b |
| IL-2 |
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IL-2 |
| IL-4 |
IL-4 |
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IL-4 |
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IL-5 |
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| IL-6 |
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IL-6 |
| IL-10 |
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IL-10 |
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IL-12p40/p70 |
| GM-CSF |
GM-CSF |
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GM-CSF |
| IFNg |
IFNg |
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IFNg |
| TNFa |
TNFa |
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TNFa |
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VEGF |
Non-Human Primate Luminex Panels
(Updated April 7, 2006)
| Bio-Rad |
Chemicon |
R&D |
Invitrogen |
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IL-2 |
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IL-2 |
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IL-4 |
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IL-10 |
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IFNg |
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IFNg |
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TNFa |
Cell Signaling Luminex Panels
(Updated April 7, 2006)
| Bio-Rad |
Chemicon |
R&D |
Invitrogen |
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Receptor Tyrosine Kinases |
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| Phospho-EGFR (*cannot be multiplexed*) |
Phospho-EGFR |
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EGFR (Tyr 1068) |
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Total EGFR |
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Phospho-PDGFRa |
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Phospho-PDGFRb (mouse) |
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Phospho-c-Kit |
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c-Kit (Tyr 823) |
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Total c-Kit |
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| Phosphorylated Proteins |
Phosphorylated Proteins |
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Phosphorylated Proteins |
| Akt1/PKBa (Ser473) |
Akt1/PKBa (Ser473) |
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Akt (Ser473) |
| ATF-2 (Thr71) |
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| Erk 1/2 (Thr202/Tyr204, Thr185/Tyr187) |
Erk/Map Kinase 1/2 (Thr185/Tyr187) |
Erk 1/2 (Trp185/Tyr187) |
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| Erk1 (Thr202/Tyr204) |
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| Erk2 (Thr185, Tyr187) |
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c-Met (Tyr 1349) |
| HSP27 (Ser78) |
HSP27 (Ser78) |
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HSP27 (Ser 82) |
| p38 MAPK (Thr180/Tyr182) |
p38/SAPK (Thr180/Tyr182) |
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p38 (Thr180/Tyr182) |
| Rsk1/MAPKAP Kinase 1a (Ser380) |
IGF1R (Tyr 1135/Tyr 1136) |
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| JNK (Thr183/Tyr185) |
JNK/SAPK1 (Thr183/Tyr185) |
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JNK (Thr183/Tyr185) |
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Lck |
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| c-Jun (cannot be plexed with ATF-2) |
c-Jun (Ser73) |
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IR (Tyr 1162/Tyr1163) |
| STAT3 (Tyr705) |
STAT3 (Tyr705) |
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STAT3 (Ser727) |
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IRS1 |
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IRS1 (Ser 312) |
| p70 S6 Kinase (Thr421/Ser424) |
p70 S6 Kinase (Thr412) |
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p70 S6 Kinase (Thr421/Ser424) |
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CREB (Ser133) |
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CREB (Ser133) |
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STAT5A/B (Tyr694/699) |
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| IkBa (Ser32/Ser36) |
IkBa (Ser32) |
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IkBa (Ser32) |
| GSK3alpha/beta (Ser21/Ser9) |
STAT6 (Tyr641) |
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GSK-3beta (Ser 9) |
| p90RSK (Thr359/Ser363) |
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PRAS (Trp 246) |
| STAT2 (Tyr689) |
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| TrkA (Tyr490) |
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| NFkB p65 (Ser 536)(cannot be plexed with IkBa) |
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| p53 (Ser 15) |
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p53 (Ser 15) |
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Tau Ser 199) |
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Tau (Thr 181) |
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STAT1 (Tyr 701) |
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RB (Thr 821) |
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RB (Ser 249/Thr 252) |
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| Total Proteins |
Total Proteins |
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Total Proteins |
| c-Jun |
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| Akt/PKB |
Akt/PKB |
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Akt |
| ATF-2 |
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HSP27 |
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HSP27 |
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Erk/MAP Kinase 1/2 |
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Erk 1/2 |
| Erk2 |
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| p38 MAPK |
p38/SAPK |
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p38 |
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Lck |
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| JNK |
JNK/SAPK1 |
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JNK |
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STAT3 |
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ß-Catenin |
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IRS1 |
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p70 S6 Kinase |
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p70 S6 Kinase |
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CREB |
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CREB |
| IkBa |
IkBa |
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IkBa |
| p90RSK |
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p53 |
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STAT1 |
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Tau |
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RB |
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Apoptosis Markers |
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Active Caspase 3 |
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Isotyping Luminex Panels
(Updated April 7, 2006)
| Bio-Rad |
Chemicon |
R&D |
Invitrogen |
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Mouse |
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IgG1 |
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IgG2a |
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IgG2b |
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IgG3 |
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IgA |
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IgM |
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IgE |
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Human |
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IgG2 |
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IgG3 |
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IgG4 |
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IgE |
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IgM |
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IgA |
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IgG |
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IgG1 |
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Price Structure
Service includes registration, processing, analyzing, deriving concentrations, graphically representing, uploading data and long term archiving of samples.
| Analysis Size |
Cost per Analyte |
Cost per Sample |
| 22-plex Analysis |
$8 |
$176 |
| 10-plex Analysis |
$15 |
$150 |
| 5-plex Analysis |
$27 |
$135 |
Data Analysis : Novel Methods of Presentation for Complex Luminex Data Sets
A significant challenge to understanding complex datasets such as microarray and Luminex is our ability to see the data. Traditional two-dimensional graphical presentations are becoming too limited as the number of analytes increases. The directors and technology development team will also be involved in the development of novel methods to easily present large amounts of complex Luminex data to investigators for the purposes of discovery and immunomonitoring. A part-time biostatistician will be extremely valuable part of developing and validating many of these presentation formats and assisting with statistical modeling. Many of the tools created for the visualization and data mining of DNA microarray data may be applicable to luminex data. BIIR Luminex core staff has already begun to develop protocols for the conversion and analysis of Luminex data by software built to analyze and present complex data sets. We are presently able to visualize the comprehensive cytokine and chemokine expression profile of large numbers of autoimmune and transplant patients undergoing therapy as part of both primary research and ongoing blinded clinical trials. At the present time we have been able to present the data as both biased and unbiased clustering of patient data, self organizing maps (SOM), multidimensional graphing, intensity mapping, and gene-tree analysis. To address some of these issues, Chemicon Invitrogen is currently developing ActivityBase compatible Luminex presentation software and has agreed to beta-test the package at the BIIR core facility. These Luminex presentation formats may prove to be powerful tools for therapeutic immunomonitoring, but could have an ever greater impact in the discovery of the underlying immune disregulation at the root of many of these diseases.
Vendor Comparison
Serum
Cell Culture Supernatants in Triplicate
ELISA Comparison
Automation of Sample Preparation by Tecan Freedom Evo 200 Robot
Many validation and correlation tests have been performed over the last few months in order to ensure us that the Tecan does not influence the Luminex assay performance. Our data suggest that the Tecan does not disrupt the assay. All lyophilized recombinant standards provided by the manufacturer's correlated well whether performed by the Tecan or manually. In addition, the Tecan shows promise in the ability to actually improve the low and high end sensitivities of the standard curves.
IL-8 Standard Curve By Hand
IL-8 Standard Curve By Tecan
Correlation
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