Flow cytometric analysis of Rsc96 cell using HB9 antibody.
Black line: Positive blank control RSC96), Negative blank control (HL60) Green line: Primary Antibody (Rabbit Anti-HLXB9 antibody (orb157435)) Orange line: Isotype Control Antibody (Rabbit IgG). Blue line: Secondary Antibody (Goat anti-rabbit IgG-AF488) RSC96 (Positive) and HL60 (Negative control) cells (black) were fixed with 4% PFA for 10 min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at -20C, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HLXB9 Antibody (orb157435) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody (blue) incubation for 40 min at room temperature. Acquisitions of 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Blank control: K562. Primary Antibody (green line): Rabbit Anti-MNX1 antibody (orb157435), Dilution: 2 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 0.5 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
Blank control: K562. Primary Antibody (green line): Rabbit Anti-MNX1 antibody (orb157435), Dilution: 2 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 0.5 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
K562 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (MNX1) polyclonal Antibody, Unconjugated (orb157435) 1:100, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
