Related topics: Alphabetical list, Secondary antibodies
On March 14, 2019
Kasper Mølgaard Jensen wrote:
AF647-F(ab’)2 antibody used in a titration experiment to stain CART19 cells expressing a chimeric antigen receptor or to stain negative PBMCs.
Ab volumes ranging from 0,5uL to 10uL were used to stain between 5-750.000 cells in remaining volume after washing (2% BSA PBS) (Figure 1). All solution and cells were keept at 4 degrees or on ice and stained in the dark for 30 minutes prior to 2x wash with 2mL 2% BSA PBS.
A stain index curve were calculated to estimate an optimal antibody volume at around 8uL for 5-750.000 cells (Figure 2).
High level of unspecific staining seen if cells were washed once, in lower volumes or if washed in plain PBS.
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