Hi, I'd like to stain 10M cells with CD3 for cell sorting. The datasheet says 5ul/test, and typically use 1 × 10^6 cells in a 100-µl experimental sample (a test). So does it mean I need a 50ul antibody for 10M cells staining? That's a lot. So anyone have experience with large-volume sample staining? What's the minimum antibody needed?
Thanks!
Hello Yao Fu,
You may not need 50ul of antibody for staining 10×10^6 cells. Cell concentration is not as important as other staining parameters. A single antibody concentration can work for a range of cell concentrations. However, certain antibodies may be sensitive to cell number.
As per the datasheet, 5ul antibody is used for 1×10^6 cells in a 100ul experimental sample. So, instead of using 50ul antibody for 10×10^6 cells in a 1ml experimental sample, you could use 25ul antibody for 10×10^6 cells in a 0.5ml experimental sample or reduce it still further to 12.5ul of antibody for 10×10^6 cells in a 0.25ml experimental sample.
Also, for comparison of results, you could run two sets in the same experiment namely, a normal set with 1×10^6 cells using 5ul of antibody in a 100ul experimental sample and a scale-up set with 10×10^6 cells using either 25ul or 12.5ul of antibody in a 0.5ml or 0.25ml experimental sample respectively.
Best.