Anti-CD13 antibody [WM15] - BSA and Azide free

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD13 antibody [WM15] - BSA and Azide free (AB252261)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD13 antibody [WM15] - BSA and Azide free (AB252261)

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).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD13 antibody [WM15] - BSA and Azide free (AB252261)

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).

• Flow Cyt

Lab

Flow Cytometry - 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 image was generated from the hybridoma version of the product.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD13 antibody [WM15] - BSA and Azide free (AB252261)

This data was developed using the same antibody clone in a different buffer formulation (ab7417). ab7417 staining CD13 in wild-type THP-1 cells (top panel) and ANPEP knockout THP-1 cells (bottom panel) (ab273759). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 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 2.5μg/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 image was generated from the hybridoma version of the product.