Mitochondrial Function Assay for KMS-11 Cells

KMS-11 cells (seeded 6 × 10⁵ viable cells in each well, 6 replicates for each group) were treated with 5μM ABT-199, 5μM AT-101, or DMSO vehicle control in RPMI medium with 10% FBS for 24 h in a manner previously described by us.4 The Seahorse XF Cell Mito Stress Test Kit (Agilent 103015-100) was used to measuring cell mitochondrial function using the Agilent Seahorse XF96 analyzer following the manufacturer’s instructions (https://www.agilent.com/cs/library/usermanuals/public/XF_Cell_Mito_Stress_Test_Kit_User_Guide.pdf) accessed on 4 April 2022). The assay uses the built-in injection ports on XF sensor cartridges to add modulators of respiration into cell well during the assay to reveal the key parameters of mitochondrial function. The modulators included in this assay kit are Oligomycin, Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP), Rotenone, and Antimycin. This figure illustrates the injection sequence of these modulators and the parameters that are obtained with this assay.

Free full text: Click here

Ailawadhi S., Parrondo R.D., Dutta N., Han B., Ciccio G., Cherukuri Y., Alegria V.R., LaPlant B.R., Roy V., Sher T., Edwards B., Lanier S., Manna A., Heslop K., Caulfield T., Maldosevic E., Storz P., Manochakian R., Asmann Y., Chanan-Khan A.A, & Paulus A. (2023). AT-101 Enhances the Antitumor Activity of Lenalidomide in Patients with Multiple Myeloma. Cancers, 15(2), 477.

Publication 2023

Seahorse XF Cell Mito Stress Test Kit Seahorse XF96 analyzer

Abt 199 Antimycin Assay Carbonyl cyanide Cell Cell function Dmso Fccp Library Mito Mitochondrial Oligomycin Phenylhydrazone Respiration Rotenone Seahorse Stress test

Corresponding Organization : Mayo Clinic in Florida

Other organizations : Mayo Clinic

Top 5 similar protocols

Variable analysis

independent variables

Treatment with 5μM ABT-199
Treatment with 5μM AT-101
Treatment with DMSO vehicle control

dependent variables

Cell mitochondrial function

control variables

Cell seeding density (6 × 10^5 viable cells per well)
Cell culture medium (RPMI with 10% FBS)
Incubation time (24 hours)

positive controls

None specified

negative controls

DMSO vehicle control

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!