Recombinant Anti-CD13 antibody [WM15] - BSA and Azide free (ab252261)

This data was developed using the same antibody clone in a different buffer formulation (ab7417)

Flow cytometry overlay histogram showing wild-type THP1 (green line) and ANPEP knockout THP1 cells (ab273759) stained with ab7417 (red line). The cells were incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab7417) (1x10⁶ in 100μl at 1 μg/ml) for 30 min at 4°C.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 4°C.

Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type THP1 cells - black line; ANPEP knockout THP1 cells ab273759 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7417).

ab7417 staining CD13 on the surface of human peripheral blood leukocytes in Flow Cytometry.

This data was developed using the same antibody clone in a different buffer formulation (ab7417). ab7417 staining CD13 in PANC-1 cells (top panel - positive control) and HEK-293 cells (bottom panel - negative control). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab7417 at 0.5μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7417).

ab7417 staining CD13 in A375 (Human epithelial cell line from skin malignant melanoma) cells.

The cells were fixed with 100% methanol (5min), permeabilized with 0.1%PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab7417 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

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

This product also produced a positive signal in A375 when fixed with 4% formaldehyde (10min).