Advanced Phenotypic Analysis

     
 
 
 
Mitochondrial Function Assays

New Probes of Mitochondrial Function

MitoPlates from Biolog provide a powerful new research tool by allowing scientists to run preconfigured sets of 96 mitochondrial function assays in one experiment. Mitochondria can be interrogated and characterized in novel ways, looking at rates of substrate metabolism, sensitivity to drugs and other chemicals, and effects of mutations in mitochondria-related genes.

Substrates or inhibitors permeate the cell membrane and enter mitochondria, stimulating or inhibiting production of NAD(P)H or FADH2 which is then measured using a tetrazolium redox dye.

Assay Principle

Investigate how mitochondria change with:

Cell differentiation

Cancer & ageing

Neurological disorders

Metabolic disorders

Immune cell activation

Bacterial/viral infection

Inborn genetic defects

Mitochondrial function is assayed by measuring the rates of electron flow into and through the electron transport chain from metabolic substrates that produce NAD(P)H or FADH2 such as L-malate, succinate, pyruvate, etc. Each substrate follows a different route, using different transporters to enter the mitochondria and different dehydrogenases to produce NAD(P)H or FADH2, The electrons travel from the beginning (complex 1 or 2) to the distal portion of the electron transport chain where a tetrazolium redox dye (MC) acts as a terminal electron acceptor that turns purple upon reduction. Additional MitoPlate assays probe the sensitivity of the mitochondria to a set of 22 diverse inhibitors.

MitoPlate S-1 with 31 Substrates:

Glycogen

D-Glucose-1-PO4

No Substrate Control

α-D-Glucose

D-Glucose-6-PO4

D-Gluconate-6-PO4

D,L-α-Glycerol-PO4

L-Lactic Acid

Pyruvic Acid

Citric Acid

D,L-Isocitric Acid

cis-Aconitic Acid

α-Keto-Glutaric Acid

Succinic Acid

Fumaric Acid

L-Malic Acid

α-Keto-Butyric Acid

D-β-Hydroxy-Butyric Acid

L-Glutamic Acid

L-Glutamine

Ala-Gln

L-Serine

L-Ornithine

Tryptamine

Sparker Malate Control

Acetyl-L-Carnitine

Octanoyl-L-Carnitine

Palmitoyl-L-Carnitine

Pyruvic Acid

γ-Amino-Butyric Acid

α-Keto-Isocaproic Acid

L-Leucine

MitoPlate I-1 with 22 Inhibitors:

No Inhibitor Control

Meclizine

No Substrate Control

Complex I Inhibitor Rotenone

Complex I Inhibitor Pyridaben

Berberine

Complex II Inhibitor Malonate

Complex II Inhibitor Carboxin

Alexidine

Complex III Inhibitor Antimycin A

Complex III Inhibitor Myxothiazol

Phenformin

Uncoupler FCCP

Uncoupler 2,4-Dinitrophenol

Diclofenac

Ionophore, K Valinomycin

Calcium / CaCl2

Celastrol

Gossypol

Nordihydro-guaiaretic acid

Trifluoperazine

Polymyxin B

Amitriptyline

Papaverine

Simple Assay Procedure:

STEP 1: Prepare and pipet assay mixture containing cell permeabilizing buffer and redox dye into appropriate wells.

STEP 2: Start the assays by adding 2x cell suspension to all wells.

STEP 3: Load the MicroPlate into the OmniLog® for kinetic reading of the rate of purple color formation.

Ordering Information:

Catalog #

Description

14104

MitoPlate I-1

14105

MitoPlate S-1

72303

Biolog MAS

74353

Biolog Redox Dye Mix MC

74354

Biolog Redox Dye Mix MD

96161

OmniLog PM-M System (NA Plug)

96162

OmniLog PM-M System (Schuko Plug)

96164

OmniLog PM-M System (UK Plug)

Not Included: Saponin permeabilizing solution and substrate solutions for

MitoPlate I-1. Please review Instructions for Use prior to ordering.

BIOLOG, Inc.

21124 Cabot Blvd Hayward, CA 94545 USA

(510) 785-2564 • www.biolog.com

Unique Features and Advantages:

MitoPlates are preloaded with 96 tests ready for use

Plates designed to measure effects of substrates and inhibitors on mitochondrial function

Easy, robust protocols with any cell type – adherent or suspension cells, transformed cell lines or primary cells

Assays need only 20,000 to 40,000 cells per well

Novel tetrazolium dye chemistry provides a terminal electron acceptor in easy to read colorimetric assays

OmniLog instrument provides automated temperature controlled incubation and kinetic reading of multiple plates (50 plates at 15 min, or 16 at 5 min intervals)

OmniLog System and Analysis Software

The OmniLog Instrument and associated software allows for real-time recording and kinetic analysis of electron flow rates. The OmniLog can simultaneously incubate and read up to 50 MitoPlates and provides powerful analysis tools to get the most from your experimental data.

Part 00A XXX, Rev A 11/2017