Oligo/FCCP/Rot/AA drug recommended conc for human melanoma cell lines (A375 and SKMEL5)

Hi Christian,

Thank you for contacting Cell Analysis Technical Support. I can see that you are already in communication with my colleague, Raji, so I will let you continue your conversation via email.

Thank you,

Courtney

Courtney Nadeau Watts

Technical Support Scientist/Remote Engineer

Cell Analysis Products

For Cell Mito stress test, Seahorse Cell Mito Stress Test Kit, the recommended working concentrations are 1.5 µM for Oligomycin and 0.5 µM for Rot/AA. The FCCP concentration needs to be optimized (0.5 µM-2 µM). The optimal final compound concentration for achieving maximal effect is cell line dependent, and may be affected by assay medium types. Therefore, it is recommended that, for each new cell line or assay medium, a titration experiment for the compounds is performed. This is especially important with FCCP, as the titration curve tends to be quite sharp, and too much FCCP can actually diminish responses in OCR. For details refer to Seahorse XF Cell Mito Stress Test Kit User Guide (agilent.com). The assays are validated with Agilent reagents, thus we cannot provide any guaranteed concentrations for in-house reagents. You may need to optimize further.

For Cell Mito stress test, Seahorse Cell Mito Stress Test Kit, the recommended working concentrations are 1.5 µM for Oligomycin and 0.5 µM for Rot/AA. The FCCP concentration needs to be optimized (0.5 µM-2 µM). The optimal final compound concentration for achieving maximal effect is cell line dependent, and may be affected by assay medium types. Therefore, it is recommended that, for each new cell line or assay medium, a titration experiment for the compounds is performed. This is especially important with FCCP, as the titration curve tends to be quite sharp, and too much FCCP can actually diminish responses in OCR. For details refer to Seahorse XF Cell Mito Stress Test Kit User Guide (agilent.com). The assays are validated with Agilent reagents, thus we cannot provide any guaranteed concentrations for in-house reagents. You may need to optimize further.