Alexa Fluor® 647 Anti-PAI1 antibody [EPR21850-82]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-PAI1 antibody [EPR21850-82] (AB303581)

ab303581 staining PAI-1 in HUVEC (Human umbilical vein epithelial cell line) cells (positive control) and HEK293 cells (negative control). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3 M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab303581 at 1 µg/ml (shown in red) and ab195883 (Alexa Fluor®488 Anti-Tubulin antibody [YOL1/34]) at 5 µg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-PAI1 antibody [EPR21850-82] (AB303581)

Flow cytometry overlay histogram showing left HUVEC positive cells and right negative HEK293 cells stained with ab303581 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab303581) (1x 106 in 100μl at 0.04μg/ml (1/12500)) for 30 min at 22°C.

Isotype control antibody (black line) was used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 40 mW Red laser (638 nm) and 660/10 bandpass filter.

This antibody gave a positive signal in HUVEC cells fixed with 80 % methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.