To stain iNKT cells specifically, mCD1d/Ig fusion proteins (CD1d dimer; mouse CD1d DimerX, BD Biosciences, San Jose, CA) were incubated overnight at 37°C with a 40-fold molar excess of α-GalCer (in PBS containing 0.5% Tween 20). The staining cocktail was prepared by mixing α-GalCer-loaded mCD1d/Ig proteins with FITC- or APC-conjugated anti-mouse IgG1 Ab (clone A85-1, BD Biosciences) at a 1 : 2 ratio of dimer to anti-mouse IgG1 Ab. Subsequently, the mixture was incubated for 2 hr at room temperature.
Fitc or apc conjugated anti mouse igg1 ab clone a85 1
Manufactured by BD
FITC- or APC-conjugated anti-mouse IgG1 Ab (clone A85-1) is a laboratory reagent used for the detection and analysis of mouse IgG1 antibodies. It consists of a mouse monoclonal antibody (clone A85-1) that is conjugated to the fluorescent dyes FITC or APC.
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2 protocols using fitc or apc conjugated anti mouse igg1 ab clone a85 1
1
Staining of iNKT Cells Using α-GalCer-Loaded CD1d Dimers
2
Staining iNKT Cells with CD1d Dimer
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To stain iNKT cells specifically, mCD1d/Ig fusion proteins (CD1d dimer; mouse CD1d dimerX, BD Biosciences) were incubated overnight at 37 °C with 40-fold molar excess of α-GalCer (in PBS containing 0.5% Tween 20). The staining cocktail was prepared by mixing α-GalCer-loaded mCD1d/Ig proteins with FITC- or APC-conjugated anti-mouse IgG1 Ab (clone A85-1, BD PharMingen, San Diego, CA) at a 1:2 ratio of dimer to anti-mouse IgG1 Ab. Subsequently, the mixture was incubated for 2 hrs at room temperature (RT).
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