Assays routinely used in our laboratory with SeedEZ and Perfused Organ Panel

Unless labeled “terminal,” all assays are mix-and-measure assays done by:
     A.  adding assay reagents to live cells, followed by collection of supernatants, or
     B.  assaying the culture medium directly for analytes consumed (e.g. glucose) or released (e.g. cytokines) by cells
Cell health can be monitored over time using method B by assaying medium when it is exchanged.
All assays are quantified using a plate reader (bioluminescence, fluorescence, or absorbance readout).
Some assays may be multiplexed and run in the same well. For example, bioluminescence and fluorescence assays.

Cell Viability and Apoptosis	Value	Method
Cell Titer Glo	ATP (terminal)	A
Cell Titer Blue	Live Cell Number (standard curve)	A
alamarBlue	Live Cell Number (standard curve)	A
CCK-8	Live Cell Number (standard curve)	A
LDH Release	Cell Death (toxicity)	B
Total LDH	Live Cell Number (terminal)	A
Live/Dead Staining	Calcein AM/ Propidium Iodide	Imaging, fluorescence microscopy
Caspase-3	Terminal	A

Functional Assays		Method
Glucose Glo	Glucose Consumption	​B
EnzyChrom Glucose	Glucose Consumption	B
L-Lactate	Lactate Production	B
QuantiChrom Urea	Urea Synthesis	B
Bile Acids	Terminal	A

Drug Metabolism		Method
P450-Glo Assays	CYP3A4, CYP1A2, CYP2C9, CYP2B6 (bioluminescence)	A
EROD (ethoxyresorufin-O-deethylation)	CYP1A Assay (fluorometric)	A
3-cyano-7-ethoxycoumarin	CYP2C19 Assay (fluorometric)	A

Inflammation		Method
Griess Assay	Nitric Oxide Production	A
TNFalpha ELISA		A
CXCL10 ELISA		A
Luminex Inflammation Panel		A

Protein		Method
Immunofluorescence	Directly in scaffold (terminal)	A
Western Blot	Media samples, or Protein extraction directly from cells in scaffold	A, B (terminal)
ELISA	Media samples, or Protein extraction directly from cells in scaffold	A, B (terminal)

Nucleic Acid		Method
DNA/RNA extraction for qPCR	Terminal	A