Anti-mouse IgG F(ab’)2 fragment specific, goat pAb, Alexa Fluor-647 conjugated, Jackson ImmunoResearch, 115-605-006

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  • 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.

    Western blotNot Rated
    ImmunostainingNot Rated
    IPNot Rated
    ChIPNot Rated
    ELISANot Rated
    Flow cytometry33333
    LuminexNot Rated
    EMNot Rated

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Western blot
Immunostaining
IP
ChIP
ELISA
Flow cytometry
Luminex
EM