Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (AB30470)

Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab30470 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30470, 1 μg/1 x 10⁶ cells) for 30 minutes at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1 μg/1 x 10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 minutes) used under the same conditions.

Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (AB30470)

Flow cytometry staining of human whole blood with ab30470 (right) or mouse IgG2aκ; (ab18413) isotype (left). Red blood cells of 200 μL blood were lysed, then cells were incubated for 30 min on ice in 1x PBS containing 10 μg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab30470) or mouse IgG2aκ; (ab18413) isotype (1x10⁶ in 100 μL at 0.2 μg/ml) for 30 min on ice.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (ab150117) was used at 1/2000 dilution for 30 min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on alive cells.