Anti-CD45 antibody [F10-89-4] - BSA and Azide free

• Flow Cyt

Lab

Flow Cytometry - Anti-CD45 antibody [F10-89-4] - BSA and Azide free (AB230296)

Flow cytometry overlay histogram showing left Jurkat positive cells and right negative HeLa stained with ab230296 (red line). The cells were incubated in 1x PBS containing 10% human IgG and 10%; normal goat serum to block non-specific protein-protein interaction followed by the antibody ab230296 at 1 μg/ml for 30min on ice.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150177) was used at 1/4000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG2a (ab18413) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on viable single cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [F10-89-4] - BSA and Azide free (AB230296)

IHC image of CD45 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab30470, 1 μg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab30470).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [F10-89-4] - BSA and Azide free (AB230296)

ab30470 staining CD45 in Jurkat (Human T cell leukemia cell line from peripheral blood) cells. The cells were fixed with 4% formaldehyde (10 minutes), then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab30470 at 5 μg/ml (shown in green) and ab206369, Rabbit monoclonal to beta Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). This was followed by an incubation at room temperature for 1 hour with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed, at 1 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in Jurkat cells fixed with 80% methanol (5 minutes).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab30470).

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD45 antibody [F10-89-4] - BSA and Azide free (AB230296)

Flow cytometry staining of human whole blood with ab230296 (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 (ab230296) 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.