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.
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 |
Lysed LDH |
Live Cell Number (terminal) |
A |
Live/Dead Staining |
Calcein AM/ Propidium Iodide |
Imaging, fluorescence microscopy |
Caspase-3 |
Terminal |
A |
CytoTox Fluor |
Cell Death (toxicity) |
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 |
Amplex Red AChE/ACh |
Acetylcholinesterase activity/Acetylcholine content |
A |
Acetylthiocholine (ATCh) and DTNB (Ellman's reagent) |
Acetylcholinesterase activity |
A |
Alanine Aminotransferase (ALT) |
ALT Enzyme Activity (terminal) |
A |
Drug Metabolism |
Method |
|
P450-Glo Assays |
CYP3A4, CYP1A2, CYP2C9, CYP2B6 (luminescent) |
A |
EROD (ethoxyresorufin-O-deethylation) |
CYP1A Assay (fluorometric) |
A |
MROD (methoxyresorufin-O-demethylation) |
CYP1A2 Assay (fluorometric) |
A |
3-cyano-7-ethoxycoumarin |
CYP2C19 Assay (fluorometric) |
A |
Stress and Inflammation |
Method |
|
Griess Assay |
RNS production (nitrite; indirect nitric oxide) |
B |
TNFalpha ELISA |
B |
|
CXCL10 ELISA |
B |
|
Luminex Inflammation Panel |
B |
|
Amplex Red |
hydrogen peroxide (indirect ROS production) |
B |
Glutathione (GSH/GSSG) |
Terminal |
A |
TBARS |
Lipid Peroxidation (terminal) |
A |
Protein |
Method |
|
Immunofluorescence |
Directly in scaffold (terminal) |
A |
Western Blot |
Media samples, or Protein extraction directly from cells in scaffold (terminal) |
A, B |
ELISA |
Media samples, or Protein extraction directly from cells in scaffold (terminal) |
A, B |
Nucleic Acid |
Method |
|
DNA/RNA extraction for qPCR |
Terminal |
A |