BioLegend
Brilliant Violet 650™ anti-human CD86 Antibody
305428
IT2.2
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The goal of this experiment was to identify CD86+ cells as a measure of dendritic cell activation.
Flow cytometry
Human dendritic cells
0.1uL Zombie Green viability stain in 100uL PBS, incubated at room temp for 15 mins
Human TruStain Fc block, 5uL diluted in 95uL FACS buffer (5% FBS in DPBS), incubated at 4°C for 10 mins
1uL of CD86 antibody diluted in 100uL of FACS buffer (5% FBS in DPBS), incubated at 4°C for 20 mins
Cytofix buffer, 100uL for 20 mins at 4°C
Flow cytometry in a beckman coulter cytoflex cytometer.
This product helped me identify true positives for CD86. I was able to identify reliable true positives, however, I did not see a true negative population. My study design involved maturing dendritic cells and stimulating activation, which may explain the absence of a true negative population, but this means that I cannot speak to the existence of such a true negative. I also saw a small spectrum of activation, resulting in several cells that I gated out as "CD86-" due to overlap with the unstained sample, when in fact they may have been somewhat positive.
N/A
Reliably identifies CD86+ true positive cells.
Somewhat activated cells likely fall in the same range as background signal.
Good signal and reliably works at 1uL concentration.