Mitochondrial Function Assay Protocol

RPMI 1640 media, fetal bovine serum, penicillin-streptomycin, Mag-Fluo-4 AM, Rhod-2 AM, Fluo-4 AM, Annexin V-FITC staining kit, Image-IT live mitochondria permeability transition pore assay kit, MitoSOX Red, MitoTracker green and DCFDA were from Thermo Fisher Scientific (Burlington, ON, Canada). L-asparaginase (ab73439) was from Abcam (Toronto, ON, Canada). 2,5-di-tert-butylhydroquinone (TBHQ) was from Sigma (Oakville, ON, Canada). Xestospongin-C (XeC), ruthenium red (RuR), and cyclosporine A (CsA) were from Bio-Techne (Oakville, ON, Canada). HAP1 (D-12) and actin (I19) antibodies, and Mito-Tempo were from Santa Cruz Biotech. (Dallas, TX, United States of America).

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Lee J.K., Rosales J.L, & Lee K.Y. (2023). Requirement for ER-mitochondria Ca2+ transfer, ROS production and mPTP formation in L-asparaginase-induced apoptosis of acute lymphoblastic leukemia cells. Frontiers in Cell and Developmental Biology, 11, 1124164.

Publication 2023

RPMI 1640 media fetal bovine serum penicillin-streptomycin Mag-Fluo-4 AM Rhod-2 AM Fluo-4 AM Annexin V-FITC staining kit Image-IT live mitochondria permeability transition pore assay kit MitoSOX Red MitoTracker green 2,5-di-tert-butylhydroquinone (TBHQ) Mito-Tempo

Actin Annexin v fitc Antibodies Assay Cyclosporine a Dcfda Di tert butylhydroquinone Fetal bovine serum Fluo 4 L asparaginase Mito Mitochondria permeability transition pore Penicillin Rhod 2 Streptomycin Xestospongin c

Corresponding Organization :

Other organizations : Alberta Children's Hospital, University of Calgary

Top 5 similar protocols

Variable analysis

independent variables

L-asparaginase (ab73439)
2,5-di-tert-butylhydroquinone (TBHQ)
Xestospongin-C (XeC)
Ruthenium red (RuR)
Cyclosporine A (CsA)
Mito-Tempo

dependent variables

Cell viability/proliferation
Mitochondrial permeability transition pore opening
Mitochondrial reactive oxygen species (ROS) production
Mitochondrial membrane potential

control variables

RPMI 1640 media
Fetal bovine serum
Penicillin-streptomycin
Mag-Fluo-4 AM
Rhod-2 AM
Fluo-4 AM
Annexin V-FITC staining kit
Image-IT live mitochondria permeability transition pore assay kit
MitoSOX Red
MitoTracker green
DCFDA
HAP1 (D-12) antibody
Actin (I19) antibody

positive controls

No positive controls specified.

negative controls

No negative controls specified.

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