The Seahorse assays were performed using the Agilent Seahorse XF Cell Mito Stress Test kit using injections of 1 μM oligomycin, 1 μM carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) and 0.5 μM each of antimycin A and rotenone at the intervals indicated. Prior to analysis, cells were equilibrated for one hour in Mito Stress medium supplemented with 1 mM pyruvate, 10 mM glucose, and 2 mM L-glutamine. The instrument is located at the Seahorse Core at Brigham and Women’s Hospital. All assays were normalized based on the number of viable cells included following drug pre-treatment to control for effects of drug treatment on viability. For assays using primary leukemia cells, cells were cultured on MS5 stroma for 72 hours prior to B-ALL cell purification by human CD45 bead enrichment (Miltenyi) followed by Seahorse Mito Stress analysis.
Human cd45 bead enrichment
Manufactured by Miltenyi Biotec
The Human CD45 bead enrichment is a laboratory product designed for the isolation and enrichment of CD45-positive cells from human samples. It utilizes magnetic beads coated with antibodies targeting the CD45 surface marker to selectively bind and separate the desired cell population. This product provides a tool for researchers to isolate and study CD45-expressing cells, which play a crucial role in various immunological processes.
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Lab products found in correlation
2 protocols using human cd45 bead enrichment
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Mitochondrial Respiration Assays in Cells
2
Mitochondrial Respiration Assays in Cells
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The Seahorse assays were performed using the Agilent Seahorse XF Cell Mito Stress Test kit using injections of 1 μM oligomycin, 1 μM carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) and 0.5 μM each of antimycin A and rotenone at the intervals indicated. Prior to analysis, cells were equilibrated for one hour in Mito Stress medium supplemented with 1 mM pyruvate, 10 mM glucose, and 2 mM L-glutamine. The instrument is located at the Seahorse Core at Brigham and Women’s Hospital. All assays were normalized based on the number of viable cells included following drug pre-treatment to control for effects of drug treatment on viability. For assays using primary leukemia cells, cells were cultured on MS5 stroma for 72 hours prior to B-ALL cell purification by human CD45 bead enrichment (Miltenyi) followed by Seahorse Mito Stress analysis.
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