Hormone & Other 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 frequency assays, denoted with an asterisk (*) below, require a sufficient number of samples to cover the cost of the assay. The minimum # of samples depends on the assay and whether the core currently has samples for that assay from other investigators.

Assay

Requirements for Human/Canine

Requirements for Mouse/Rat

*ACTH (RIA)

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

55 ul plasma for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The assay range is 5-100 pg/ml with 300 ul (so with 55 ul the detection range is 27-545 pg/ml). 5-day double antibody assay.

Amino Acids - Full and Gluconeogenic profiles - (HPLC)

100 ul plasma or supernatant (not urine) from tissue prep. Detection range: >1 umol/l.

50 ul plasma or supernatant (not urine) from tissue prep. Detection range: >1 umol/l.

Catecholamines - Epinephrine and Norepinephrine (HPLC)

EPI & NE are measured together from 250 ul plasma or supernatant (not urine) from tissue prep. EGTA-glutathione must be added at the time of collection. 1-day assay. Detection range: 25-1000 pg/ml (a smaller extraction volume can be used to measure >1000 pg/ml). Store samples at -70°C.

EPI & NE are measured together from 50 ul plasma or supernatant (not urine) from tissue prep. EGTA-glutathione must be added at the time of collection. 1-day assay. Detection range: 25-1000 pg/ml (a smaller extraction volume can be used to measure >1000 pg/ml). Store samples at -70°C.

*Corticosterone (RIA)

NA

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

Cortisol - plasma (RIA)

100 ul plasma for duplicate analysis. 2-day double antibody assay. Detection range: 0.5-100 ug/dl.

NA

Cortisol - salivary (RIA)

0.5 ml saliva for duplicate analysis. 2-day double antibody assay. Detection range: 1-20 ng/ml. Saliva collection procedures.

NA

*C-peptide (RIA) - human & canine)

400 ul plasma for duplicate analysis. Needs EDTA/Trasylol added at the time of collection. Regular assay: 2-day double antibody, detection range 0.1-5 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.

*Ghrelin - Active (RIA)

400 ul 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 ul plasma for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The assay range is 5-100 pg/ml with 300 ul (so with 30 ul the detection range is 50-1000 pg/ml). 5-day double antibody assay. Needs Pefabloc SC (AEBSF) added at the time of collection.

*Ghrelin - Total (RIA)

400 ul plasma for duplicate analysis. 2-day double antibody assay. Detection range: 150-5000 pg/ml.

30-60 ul plasma for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The assay range is
100-1000 pg/ml with 300 ul (so with 30 ul the detection range is 1000-10,000 pg/ml). 5-day double antibody assay.

*Glucagon (ELISA)

25 or 60 ul 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, GLP-2 or GRPP. Detection range 2-180 or 1.5-130 pmol/l (5-453 pg/ml), respectively.

25 or 60 ul 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, GLP-2 or GRPP. Detection range 2-180 or 1.5-130 pmol/l (5-453 pg/ml), respectively.

Glucagon (RIA)

400 ul of plasma for duplicate analysis. Needs EDTA/Trasylol added at the time of collection. 3-day double antibody assay. Detection range: 25-200 pg/ml. Divide pg/ml by 3.45 to convert to pmol/l.

Available as an ELISA.

Glucose (enzymatic)

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

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

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

400 ul plasma for duplicate analysis. 2-day double antibody assay. Detection range: 2-150 uU/ml. Multiply uU/ml by 6 to convert to pmol/l.

30-100 ul plasma for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The undiluted assay range is 0.01-0.5 ng/ml (sensitive assay) and 0.1-10 ng/ml regular assay) with 300 ul. So, if 30 ul is provided, the assay becomes 10x less sensitive, i.e. the assay will have a detection range of 0.1-5 ng/ml for the sensitive assay and 1-100 for the regular assay. 5-day double antibody assay. Interspecies and human-specific assays are available. 1 uU/ml = 0.04 ng/ml; 1 uU/ml = 6 pmol/l.

*Leptin (RIA)

400 ul plasma for duplicate analysis. 2-day double antibody assay. Detection range: 0.5-100 ng/ml. 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.

***A minimum number of samples is required to cover our costs (typically at least 27). Contact the core for more information.***

Most Frequently Used:

  • Human Cytokine/Chemokine Panel 1 [60 ul - 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 ul - Amylin (active)*, Amylin (total)*, 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. Active and total amylin, and active and total GLP-1 cannot be run together in the same assay.

  • Human Metabolic Hormone Panel V3 [60 ul - Amylin (active)*, Amylin (total)*, C-Peptide, Ghrelin (active)*, GIP, GLP-1(active)*, GLP-1 (total)*, Glucagon*, IL-6, Insulin, Leptin, MCP-1, Pancreatic Polypeptide, PYY, TNFa, Secretin]. *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. Active and total amylin, and active and total GLP-1 cannot be run together in the same assay.

  • Human Th17 Panel [60 ul - 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 Cytokine/Chemokine Panel 1 [5-60 ul 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 Th17 Panel [60 ul - 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 [30 ul - 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.

 

 

 

 

 

 

 

 

 

 

 

PYY-Active (3-36) (RIA)

300 ul 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 ul 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 ul plasma for duplicate analysis. 2-day double antibody assay. Detection range: 0.1-10 ng/ml.

25 ul (male) or 35 ul (female) for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The assay range is 0.005-1 ng/ml with 300 ul (so with 25 ul the detection range is 0.06-12 ng/ml). 2-day double antibody assay.

Testosterone - salivary (RIA)

0.5 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 ul plasma for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The assay range is 0.05-5 ng/ml with 300 ul (so with 30 ul the detection range is 0.5-50 ng/ml). 5-day double antibody assay.

*T4 (RIA)

NA

30 ul plasma for duplicate analysis is typical, but the required sample volume depends on the hormone concentration and dilution factor. The assay range is 0.5-20 ng/ml with 300 ul (so with 30 ul the detection range is 5-200 ng/ml). 5-day double antibody assay.

*Low frequency assays require a sufficient number of samples to cover the cost of the assay. The minimum # of samples depends on the assay and whether the core currently has samples for that assay from other investigators.

 

***This core is supported by NIH grant DK020593 (DRTC). Please include an acknowledgment in your publications.***