Analytical Assays

These assays are employed to qualitatively assess or quantify the presence, amount, or functional activity of a target entity.

Immunoassays

Immunoassays are bioanalytical methods used in drug discovery that use the specificity of an antigen-antibody reaction to detect and quantify target molecules in biological samples.

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Luminex Multiplex

xMAP Technology has application in serological assays by easy detection of antibodies. A serology assay enables the detection of an antibody (target) with the use of a capture protein antigen attached to the surface of a microsphere. A detection antibody that incorporates a fluorescent label is used to quantify the amount of target antibody present. This assay is useful when serum antibodies need to be measured, such as for the response to a vaccine, allergic reactions, or autoantibody detection.

It combines the efficiency and sensitivity of traditional ELISAs with high through-put capacity, small sample volume, efficiency in terms of time and cost,high reproducibility and sensitivity and analyte quantification over a wide range of concentrations.

For more information on how Cellomatics Biosciences can assist with your assays, please contact us or request a consultation.

Applications:

  • Measurement of cytokine/chemokine/inflammatory mediator expression in a variety of biological specimens which can be applied for biomarker discovery and Target validation
  • Allergy testing
  • Single nucleotide polymorphisms (SNPs) testing
  • Diagnosis of infectious diseases
  • Application areas include inflammation, autoimmune diseases, asthma, obesity/diabetes, neurological disorders, cancer, cardiovascular disease, drug discovery and vaccine development

Biomarker qualification and validation:

  • Robust validated technology
  • Simultaneous detection and quantification of multiple analytes in a single sample from a variety of biological specimens
  • High sensitivity and reproducibility
  • High through-put capacity
  • Efficiency in terms of time and cost
  • Various species

Multiplex Assay (xMAP)

Picture: Methods- ThermoFisher Scientific & R&D Systems®; Results- Cellomatics BioSciences Ltd.

Multiplex Analysis-Serum

Serum samples from 9 healthy volunteers were analysed for GDF1, Adam TS13 and sVAM-1 using Luminex Multiplex assay. Standard curves and median fluorescent intensities (MFIs) are extracted from the Luminex MAGPix instrument. The Exponent software (Luminex) analyses the MFIs for each sample and concentrations are calculated from the standard curves. These concentrations are then graphed on GraphPad Prism.

NoPlexAssay PanelDescription
141-plexHuman Cytokine/Chemokine Panel IsCD40L, EGF, Eotaxin/CCL11, FGF-2, Flt-3 ligand, Fractalkine, G-CSF, GM-CSF, GRO, IFN-α2, IFN-γ, IL-1α, IL-1β, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17, IP-10, MCP-1, MCP-3, MDC (CCL22), MIP-1α, MIP-1β, PDGF-AA, PDGF-AB/BB, RANTES, TGF-α, TNF-α, TNF-β, VEGF
229-plexHuman Cytokine/Chemokine Panel IIEGF, G-CSF, GM-CSF, IFN-α2, IFN-γ, IL-1α, IL-1β, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17, IP-10, MCP-1, MIP-1α, MIP-1β, TNF-α, TNF-β, VEGF, Eotaxin/CCL11
323-plexHuman Cytokine/Chemokine Panel III6Ckine, BCA-1, CTACK, ENA-78, Eotaxin-2, Eotaxin-3, I-309, IL-16, IL-20, IL-21, IL-23, IL-28A, IL-33, LIF, MCP-2, MCP-4, MIP-1d, SCF, SDF-1A+β, TARC, TPO, TRAIL, TSLP
425-plexHuman Th17 PanelGM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17A, IL-17F, IL-17E/IL-25, IL-21, IL-22, IL-23, IL-27, IL-28A, IL-31, IL-33, MIP-3α/CCL20, TNF-α, TNFβ
55-plexHuman Diabetes PanelInsulin, Glucagon, Leptin, GLP-1 (Active), C-Peptide
611-plexHuman Kidney Injury PanelCalbindin, Collagen IV, FABP1, GSTα, GSTπ, IP-10, KIM-1, Osteoactivin, Renin, TFF-3, TIMP-1
78-plexHuman Src Family Kinase Panel – PhosphoproteinBlk (Tyr389), Fgr (Tyr412), Fyn (Tyr420), Hck (Tyr411), Lck (Tyr394), Lyn (Tyr397), Src (Tyr419), Yes (Tyr421)
817-plexHuman Angiogenesis/Growth Factor PanelAngiopoietin-2, BMP-9, EGF, Endoglin, Endothelin-1, FGF-1, FGF-2, Follistatin, G-CSF, HB-EGF, HGF, IL-8, Leptin, PLGF, VEGF-A, VEGF-C, VEGF-D
97-plexDNA Damage/Genotoxicity PanelATR (Total), Chk1 (Ser345), Chk2 (Thr68), H2A.X (Ser139), MDM2 (total), p21 (Total), p53 (Ser15)
106-plexHuman Apolipoprotein PanelApo AI, Apo AII, Apo B, Apo CII, Apo CIII, Apo E
1112-plexHuman Bone Metabolism PanelACTH, DKK-1, FGF-23, IL-1β, IL-6, Insulin, Leptin, Osteocalcin, OPN – Osteopontin, Osteoprotegerin, PTH, SOST, TNF-α
129-plexHuman Cancer/ Metastasis Biomarker PanelDKK-1, GDF-15, NSE, OPG, Osteonectin (SPARC), Periostin, TRAP5, TWEAK, YKL40
1310-plexHuman Cardiovascular Disease (CVD) Panel 1ADAMTS13, D-Dimer, GDF-15, Myoglobin, sICAM-1, MPO, P-selectin, Lipocalin-2/NGAL, sVCAM-1, SAA.
1414-plexHuman Cardiovascular Disease (Acute Phase) Panel 2AGP, Haptoglobin, SAP, Adipsin,  Human a2, CRP, Fetuin, Fibrinogen, L-Selectin, PF4, vonWillebrand Factor
1524-plexHuman Circulating Cancer Biomarker Panel IAFP, CEA, CYFRA21-1, FGF-2, HE4, PSA Total, PSA free, HGF, IL-6, IL-8, Leptin, MIF, OPN – Osteopontin, Prolactin, SCF, sFas, sFasL, TGF-α, TNF-α, TRAIL, VEGF, CA125, CA15-3, CA19-9, β-HCG
165-plexHuman Circulating Cancer Biomarker Panel 2Antithrombin III, Complement Factor H, Extracellular Matrix Protein 1, Vitamin D Binding Protein, Vitronectin
177-plexHuman Early Apoptosis PanelAkt (Ser473), BAD (Ser112), Bcl-2 (Ser70), Active Caspase 8 (Asp384), Active Caspase 9 (Asp315), JNK (Thr183/Tyr185), p53 (Ser46)
1811-plexHuman Kidney Injury Panel 1Calbindin, Collagen IV, FABP1, GSTα, GSTπ, IP-10, KIM-1, Osteoactivin, Renin, TFF-3, TIMP-1
197-plexHuman Kidney Injury Panel 2α-1-Microglobulin, Albumin, Clusterin, Cystatin C, EGF, Lipocalin-2/NGAL, Osteopontin (OPN)
2113-plexHuman Metabolic Hormone PanelAmylin (Total), Amylin (Active), C-Peptide, Ghrelin, GIP, GLP-1 (Active), GLP-1 (Total), Glucagon, IL-6, Insulin, Leptin, MCP-1, PP, PYY, TNF-α
209-plexHuman Liver Protein Panel – MetabolismAFP, ANGPTL3, ANGPTL4, ANGPTL6/AGF, FABP1, FGF-19, FGF-21, FGF-23, HGF
2113-plexHuman Metabolic Hormone PanelAmylin (Total), Amylin (Active), C-Peptide, Ghrelin, GIP, GLP-1 (Active), GLP-1 (Total), Glucagon, IL-6, Insulin, Leptin, MCP-1, PP, PYY, TNF-α
2214-plexHuman Soluble Cytokine Receptor PanelsCD30, sEGFR, sgp130, sIL-1RI, sIL-1RII, sIL-2Rα, sIL-4R, sIL-6R, sRAGE, sTNFRI, sTNFRII, sVEGF-R1, sVEGF-R2, sVEGF-R3
237-plexT-Cell Receptor Signaling PanelCD3ε (pan Tyr), CREB (S133), ERK (Thr185/Tyr187), LAT (pan Tyr), Lck (pan Tyr), Syk (pan Tyr), ZAP-70 (pan Tyr)
245-plexHuman Neurodegenerative Disease PanelHuman Neurodegenerative Disease Panel
253-plexHuman Sepsis PanelFGF-13, Olfactomedin-4 (OLFM-4), MMP-8
267-plexHuman Skin Immune Response PanelCortisol, HSA, Fibronectin, LPS, Involucrin, Keratin-6, Keratin-1,10
2713-plexHuman High Sensitivity T Cell PanelGM-CSF, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, TNF-α
287-plexTotal Mitogenesis RTK PanelEGFR (Total), ErbB2/HER2 (Total), ErbB3/HER3 (Total), ErbB4/HER4 (Total), IR (total), c-Met/HGFR (Total), IGF-1R (Total)
299-plexMulti-Pathway Total Cell Signaling PanelErk/MAPK 1/2 (Total), Akt (Total), STAT3 (Total), JNK (Total), p70 S6 Kinase (Total), NFκB (Total), STAT5A/B (Total), CREB (Total), p38 (Total)
305-plexHuman Oxidative Stress Magnetic Bead Panel – Cellular Metabolism Multiplex AssayCatalase, PRX2, SOD1, SOD2, TRX1

ELISA

ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying soluble substances such as peptides, proteins, antibodies, and hormones.

Flow Cytometry (FACS)

Flow cytometry is a laser-based technology used to analyse the characteristics of cells or particles. It is a widely used method for analysing the expression of cell surface and intracellular molecules, characterising and defining different cell types in a heterogeneous cell population, assessing the purity of isolated subpopulations, and analysing cell size and volume. It allows simultaneous multi-parameter analysis of single cells.

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Neutrophil characterisation

Neutrophils were isolated from fresh peripheral blood using commercially available isolation kit and the purity of cells analysed by FACS for neutrophil surface markers (CD45 and CD66b). Figure A.1-4 represents the gating used for characterising the neutrophils. The neutrophil population was stimulated with TNF-a for 4 hours. A difference in the LFA-1 surface antigen expression in unstimulated (B.1) and stimulated (B.2) neutrophil was observed.

THP-1 characterisation after PMA-differentiation

THP-1 (a model for human monocytes cells) on stimulation with PMA, differentiate and show macrophage-like phenotypes. The PMA-differentiated THP-1 cells were stained with macrophage surface markers (CD11; CD40) and analysed using FACS. From Figures A and B, it was observed that the expression of CD11 and CD40 was higher in PMA-differentiated THP-1 cells (red) when compared to undifferentiated THP-1 (green) cells.

Western Blotting

This is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or cell extracts.

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Western blotting – near IR imaging

Near IR imaging of anti-RIP kinase protein conducted on cell lysates separated on a 8% SDS-PAGE gel.

Western blotting – near IR imaging

Human Primary Epithelial Cells were transfected with the following ON TARGETplus smart pool reagents: non-targeting control (siCTRL) and siACTB using Lipofectamine.
A] Near IR imaging of Human Primary Epithelial Cell lysates captured 24 hours post-transfection, after SDS-PAGE separation and immunoblotting using anti-β-actin and anti-GAPDH antibodies.
B] 3D densitometry calculated using the Q9 Alliance© Software, for figures reported in A]. Analysis confirmed reduction of β-actin band in cells transfected with the siACTB, when compared to controls (rVI=relative volume intensity).

Request a consultation with Cellomatics Biosciences today

Our experienced team of in vitro laboratory scientists will work with you to understand your analytical assay development needs and provide a bespoke project plan with a professional, flexible service and a fast turnaround time.

To request a consultation where we can discuss your exact requirements, please contact Cellomatics Biosciences.