Oligomycin is not working

Hi Chanyoung,

Thank you for contacting Cell Analysis Technical Support. We recommend using 1.5uM oligo concentration for most cell lines (when using our kits). Are you using an Agilent kit? If you would like, I can take a look at your data. You can send it to cellanalysis.support@agilent.com. 

Thank you!

Courtney 

Courtney Nadeau Watts

Technical Support Scientist/Remote Engineer

Cell Analysis Products

Hi, Courtney Watts

I have just sent my experimental data via email with the subject line "Data for the question 'Oligomycin is not working.'"

Instead of using a kit, I used the product purchased from Sigma for Oligomycin.

I have another question.
I observed another researcher adding 1% BSA (free fatty acid) to the analysis buffer for OCR measurement experiments. However, the Agilent protocol did not include BSA. I am curious about how this might affect the experiment.

Thank you for your time and consideration.

Hi, Courtney Watts

I have just sent my experimental data via email with the subject line "Data for the question 'Oligomycin is not working.'"

Instead of using a kit, I used the product purchased from Sigma for Oligomycin.

I have another question.
I observed another researcher adding 1% BSA (free fatty acid) to the analysis buffer for OCR measurement experiments. However, the Agilent protocol did not include BSA. I am curious about how this might affect the experiment.

Thank you for your time and consideration.

Hi Chanyoung,

I will look out for your results files to see if I can see anything technically wrong in your data. However, we cannot troubleshoot reagents from another vendor as we cannot speak to the quality of the products.  

Regarding BSA: some cells/applications do need BSA during the assay (example - XF Palmitate Oxidation Stress Test - Advanced Assay where BSA is needed to keep palmitate in solution). If this is necessary, BSA containing solutions should not be loaded into the ports as this will cause issues with injections. Additionally, if there is BSA present, FCCP will need to be re-titrated as BSA will bind to FCCP, reducing its efficacy. 

Thank you,

Courtney

Hi, Courtney Watts

Thank you for your kind response.

I have sent the necessary data again via email.

Please check it.