Performance comparisonSelected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
Flow cytometric comparison of different clones for HLA-A3. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis from HLA-A3+ und HLA-A3– donors were stained with HLA-A3 antibodies and with a suitable counterstaining. As a control, HLA-A3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation dataTo provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
Comparison of staining pattern on non-fixed and fixed cells using HLA-A3 (REA950). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
