All REAfinity™ Antibodies contain a specifically mutated human IgG1 Fc region that abolishes their binding to Fcγ receptors. This allows for background-free analysis and eliminates the need for additional blocking steps such as using a FcR blocking reagent.
Human PBMCs were stained with either a mouse monoclonal CD158a-PE antibody or REAfinity CD158a-PE (clone:REA284). Staining was performed without (top) and with (bottom) FcR block. The mouse monoclonal antibody binds unspecifically to CD56-negative cells (A). Staining with the REAfinity Antibody, shows no background signal. Cells were analyzed by flow cytometry on the MACSQuant® Analyzer 10. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide (PI) fluorescence.
Staining protocols for immunophenotyping of human and mouse surface markers can be significantly faster with REAfinity Antibodies.
Steps: | Time: |
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Prepare 106 cells | |
Add blocking reagent to cells and incubate at 4°C or RT | 10–15 min. |
Add primary antibodies and incubate at 4°C or RT | 10–30 min. |
Wash cells | 1–2 x 10 min. |
Resuspend and analyze cells | |
Total staining procedure | 30–65 min. |
Steps: | Time: |
---|---|
Prepare 106 cells | |
Add blocking reagent to cells and incubate at 4°C or RT | – |
Add primary antibodies and incubate at 4°C or RT | 10 min. |
Wash cells | 10 min. |
Resuspend and analyze cells | |
Total staining procedure | 20 min. |