Assays routinely used in our laboratory with SeedEZ and PerfusionPal

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.  adding assay reagents to media taken from live cultures (e.g. after media changes)
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		​B
EnzyChrom Glucose		B
L-Lactate		B
QuantiChrom Urea		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