BD Horizon™ BV421 Goat Anti-Rabbit IgG

Immunofluorescent Staining using BD Horizon™ BV421 Goat Anti-Rabbit Ig     Left Panel - Immunofluorescence staining of apoptotic cells. HeLa cells were treated with Camptothecin (20 µM, 6 hr) to induce apoptosis. Cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized (5 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050), washed with 1× PBS, and blocked (30 min) with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1× PBS. The cells were stained (1 hr) with Purified Mouse Anti-Cytochrome c (Cat. No. 556432) and Purified Rabbit Anti-Active Caspase-3 (Cat. No. 559565) antibodies at 2.5 µg/mL diluted in blocking buffer. After washing, the cells were stained (45 min) with the second step reagents, BD Horizon™ BV421 Goat Anti-Rabbit Ig (Cat. No. 565014; pseudo-colored green) and BD Horizon™ BV480 Goat Anti-Mouse Ig (Cat. No. 564877, pseudo-colored red) antibodies at 2.5 µg/mL diluted in blocking buffer. DRAQ5 (Cat. No. 564902/564903; pseudo-colored blue) was used as a nuclear counterstain. Images were acquired with a standard epifluorescence microscope with a 20× objective.     Right Panel - Multiparameter flow cytometric analysis of CD88 expression on human peripheral blood leucocytes. Whole blood was either not stained with a primary antibody as a control (Top Plot) or primarily stained with Purified Rabbit Anti-Human CD88 monoclonal antibody (Cat. No. 559159; Bottom Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The cells were washed and secondarily stained with BD Horizon™ BV421 Goat Anti-Rabbit Ig (Cat. No. 565014). Two-parameter flow cytometric contour plots showing the correlated expression of CD88 (or Unstained control) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.