Assays

If you expect a low diluted sample concentration you may request to have it run in singlicate instead of duplicate to double the sample's concentration in the assay.

Note: Low use assays require a minimum # of samples before the assay can be performed - denoted with an asterisk below (*) 

Assay Requirements for Human/Canine Requirements for Mouse/Rat

*ACTH (RIA)

400 µl plasma for duplicate analysis. 5-day double antibody assay. Detection range: 5-100 pg/ml. 55 µl plasma for duplicate analysis. 5-day double antibody assay. Detection range: 5-100 pg/ml.

Amino Acids - full and gluconeogenic profiles - (HPLC)

100 µl plasma. Detection range: >1 µmol/l. 50 µl plasma. Detection range: >1 µmol/l.

Catecholamines – Epinephrine and Norepinephrine (HPLC)

250 µl plasma. Needs EGTA-glutathione added at the time of collection (if not added see FAQ). 1-day assay. Detection range: 50-2000 pg/ml. Store samples at -70°C. 50 µl plasma. Needs EGTA-glutathione added at the time of collection (if not added see FAQ). 1-day assay. Detection range: 50-2000 pg/ml. Store samples at -70°C.

*Corticosterone (RIA)

NA 15 µl plasma for duplicate analysis. 1-day double antibody assay. Detection range: 25-1000 ng/ml.

Cortisol - Plasma (RIA)

100 µl plasma for duplicate analysis. 1-day coated tube assay. Detection range: 0.5-50 µg/dl. NA

Cortisol - Salivary (RIA)

1 ml saliva for duplicate analysis. 5-day double antibody assay. Detection range: 0.5-20 ng/ml. Saliva collection procedures. NA

C-peptide  (RIA - human & canine)

400 µl plasma for duplicate analysis. Needs EDTA/Trasylol added at the time of collection.

Regular assay: 2-day double antibody, detection range 0.2-5 ng/ml (human) and 0.1-10 ng/ml (canine).

Sensitive assay: 5-day double antibody, detection range 0.05-2 ng/ml (human) and 0.02-0.7 ng/ml (canine).

Available as a Luminex assay. Needs EDTA/Trasylol added at the time of collection.

Creatinine (HPLC)

NA 30 µl plasma minimum

*Ghrelin - Active (RIA)

400 µl plasma for duplicate analysis. Needs Pefabloc SC (AEBSF) added at the time of collection. 2-day double antibody assay. Detection range: 8-250 pg/ml. 30-60 µl plasma for duplicate analysis. Needs Pefabloc SC (AEBSF) added at the time of collection. 5-day double antibody assay. Detection range: 5-100 pg/ml.

*Ghrelin - Total (RIA)

400 µl plasma for duplicate analysis. 2-day double antibody assay. Detection range: 150-5000 pg/ml. 30-60 µl plasma for duplicate analysis. 5-day double antibody assay. Detection range: 100-1000 pg/ml.

*Glucagon (ELISA-10 or ELISA-25)

25 or 60 µl plasma for duplicate analysis, depending on the assay. Improved sensitivity and specificity and smaller sample volume compared to RIA. *Requires a minimum # of samples before it can be performed. Little or no cross-reactivity to peptides oxyntomodulin, glicentin, mini-glucagon, GLP-1, GLP2 or GRPP. Detection range 2-180 or 1.5-130 pmol/l (5-453 pg/ml), respectively. 25 or 60 µl plasma for duplicate analysis, depending on the assay. Improved sensitivity and specificity and smaller sample volume compared to RIA. *Requires a minimum # of samples before it can be performed. Little or no cross-reactivity to peptides oxyntomodulin, glicentin, mini-glucagon, GLP-1, GLP2 or GRPP. Detection range 2-180 or 1.5-130 pmol/l (5-453 pg/ml), respectively.

Glucagon (RIA)

400 µl of plasma for duplicate analysis. Needs EDTA/Trasylol added at the time of collection. 3-day double antibody assay. Detection range: 30-400 pg/ml. Divide pg/ml by 3.45 to convert to pmol/l. 60 µl plasma for samples with basal glucagon concentrations (~50 pg/ml). 30 µl plasma for concentrations >100 pg/ml. Duplicate analysis in plasma preserved in EDTA/Trasylol at the time of collection. 5-day double antibody assay. Detection range: 15-200 pg/ml. Divide pg/ml by 3.45 to convert to pmol/l.

Glucose (Enzymatic)

100 µl of plasma for duplicate analysis. Detection range: 20-900 mg/dl. 40 µl of plasma for duplicate analysis. Detection range: 20-900 mg/dl.

Growth Hormone (RIA-human)

400 µl plasma for duplicate analysis.1-day assay. Detection range: 0.5-50 ng/ml. NA

Insulin (RIA: human & canine, mouse & rat)

400 µl plasma for duplicate analysis. 2-day double antibody assay. Detection range: 1-100 µU/ml. Multiply µU/ml by 6 to convert to pmol/l. 30 µl plasma for duplicate analysis. 5-day double antibody assay w/dilution procedure. Detection range: 0.01- 0.5 or 0.1-10 ng/ml. Interspecies or human-specific assays are available.

*Leptin (RIA)

400 µl plasma for duplicate analysis. 2-day double antibody assay. Detection range: 0.5-100 ng/ml. FOR HUMAN ONLY - No canine assay available. Available as a Luminex assay

Luminex Assays (Fluorescence)

All analytes within a panel can be measured simultaneously from a single sample. Contact us for help with your kit selection.

See here for additional information about Luminex assays.

Other assays may be available upon request.

Most Frequently Used:

  • Human Adipokine Panel 1 [20 µl - Adiponectin (total), Resistin, PAI-1 (total), Lipocalin-2/NGAL, Adipsin].
  • Human Cytokine/Chemokine Panel 1 [60 µl - EGF, Eotaxin/CCL11, FGF-2, Flt-3 Ligand, Fractalkine/CX3CL1, G-CSF, GM-CSF, GRO, IFNa2, IFN-y, IL-1a, IL-1b, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8CXCL8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1/CCL2/CXDL10, MCP-3/CCL7, MDC/CCL22, MIP-1a/CCL3, MIP-1b/CCL4, PDGF-AA, PDGF-AB/BB, RANTES/CCL5, sCD40L, TGFa, TNFa, TNFb/LTA, VEGF].
  • Human High Sensitivity T Cell [60ul - Fractalkine, GM-CSF, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IL-17A, IL-21, IL-23, ITAC, MIP-1α, MIP-1β, MIP-3α, TNF-α].

  • Human Metabolic Hormone Panel [60 µl - Active Amylin*, Total Amylin*, C-Peptide, Ghrelin (active)*, GIP (total), GLP-1(active)*, GLP-1 (total)*, Glucagon*, IL-6, Insulin, Leptin, MCP-1/CCL2, Pancreatic Polypeptide, PYY (total), TNFa]. *Needs preservative: after collecting blood sample, invert tube several times to mix, immediately add DPPIV inhibitor (for GLP-1 measurement), protease inhibitor cocktail (for Amylin measurement), Aprotinin (for Glucagon measurement) and serine protease inhibitor (for Active Ghrelin measurement). Millipore recommends using Millipore's DPP-IV inhibitor (Cat # DPP4), Sigma’s protease inhibitor cocktail, and Roche’s Pefabloc SC (AEBSF), following the manufacturers’ instructions. 
  • Human IGF Panel [60 µl - IGF-1, IGF-2].
  • Human Th17 Panel [60 µl - GM-CSF, INFy, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12(p70), IL-13, IL-15, IL-17A, IL-17E/IL-25, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-28A, IL-31, IL-33, MIP-3a/CCL20, TNFa, TNFb/LTA].

Most Frequently Used:

  • Mouse Single Plex Adiponectin Panel [7 µl - Adiponectin (total)].
  • Mouse Cytokine/Chemokine Panel 1 [5-60 µl depending on dilution - Eotaxin/CCL11, G-CSF, GM-CSF, IFN-y, IL-1a, IL-1b, 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, IP-10/CXCL10, KC/CXCL1, LIF, LIX, MCP-1/CCL2, M-CSF, MIG/CXCL9, MIP-1a/CCL3, MIP-1b/CCL4, MIP-2/CCL8, RANTES/CCL5, TNFa, VEGF].
  • Mouse/Rat IGF-1 Panel [10 µl - IGF-1].
  • Mouse Th17 Panel [60 µl - GM-CSF, INFy, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12(p70), IL-13, IL-15, IL-17A, IL-17E/IL-25, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-28A, IL-28B, IL-31, IL-33, MIP-3a/CCL20, TNFa, TNFb/LTA].
  • Mouse High Sensitivity T Cell [60ul - GM-CSF, IFNγ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12 (p70), IL-13, IL-17A, KC/CXCL1, LIX, MCP-1, MIP-2, TNF-α].

  • Mouse Metabolic Hormone Panel - note warning below - [30 µl - Amylin (Active)*, C-Peptide 2, Ghrelin (Active)*, GIP (Total), GLP-1 (Active)*, Glucagon*, IL-6, Insulin, Leptin, MCP-1, PP, PYY, Resistin, TNF-α]. We have had poor results measuring GLP-1 and glucagon (low sensitivity) with this panel, and do not recommend it for that purpose. Leptin is typically measurable but there is often poor precision. *Needs preservative: after collecting blood sample, invert tube several times to mix, immediately add DPPIV inhibitor (for GLP-1 measurement), protease inhibitor cocktail (for Amylin measurement), Aprotinin (for Glucagon measurement) and serine protease inhibitor (for Active Ghrelin measurement). Millipore recommends using Millipore’s DPP-IV inhibitor (Cat # DPP4), Sigma’s protease inhibitor cocktail, and Roche’s Pefabloc SC (AEBSF) following manufacturers’ instructions.

Purine Nucleotides (HPLC)

In situ collected and weighed, 100 mg of liver (liquid nitrogen), add 1 ml cold 0.4M perchloric acid with 0.5mM EGTA, homogenize, sit 1 min on ice, spin at ~3200g @4C for 5 min, decant supernatant, add 340ul 0.5M K2CO3, mix, sit 5 min on ice, spin ~3200g @4C for 5 min, decant, save supernatant, freeze / analyze.

In situ collected and weighed, 100 mg of liver (liquid nitrogen), add 1 ml cold 0.4M perchloric acid with 0.5mM EGTA, homogenize, sit 1 min on ice, spin at ~3200g @4C for 5 min, decant supernatant, add 340ul 0.5M K2CO3, mix, sit 5 min on ice, spin ~3200g @4C for 5 min, decant, save supernatant, freeze / analyze.

PYY-Active (3-36) (RIA)

300 µl plasma for duplicate analysis. Needs EDTA/Trasylol & DPP-IV inhibitor added at the time of collection. Detection range: 29-931 pg/ml. Available as a Luminex assay.

PYY-Total (RIA)

300 µl plasma for duplicate analysis. Needs EDTA/Trasylol added at the time of collection. Detection range: 29-931 pg/ml. Available as a Luminex assay.

*Testosterone - plasma (RIA)

400 µl plasma for duplicate analysis. 2-day double antibody assay. Detection range: 0.1-10 ng/ml. 25 µl (male) or 35 µl (female) plasma for duplicate analysis. 2-day double antibody assay. Detection range: 0.005-1 ng/ml.

Testosterone - salivary (RIA)

1 ml saliva for duplicate analysis. 2-day double antibody assay. Detection range: 0.005-1 ng/ml. Saliva collection procedures. NA

*T3 (RIA)

NA 30-60 µl plasma (depending on the animal) for duplicate analysis. 5-day double antibody assay. Detection range: 0.05-5 ng/ml.

*T4 (RIA)

NA 30 µl plasma for duplicate analysis. 5-day double antibody assay. Detection range: 0.5-20 ng/ml.

*Low use assays require a minimum # of samples before the assay can be performed.

Lipids are measured by the VUMC Lipid Core

 

***This core is supported by NIH grants DK059637 (MMPC) and DK020593 (DRTC). Please include an acknowledgment in your publications.***