PAI1 Mouse Monoclonal Antibody [A9G8]

☑ Cell treatment (CT)

Western blot analysis of PAI1 on different lysates with Mouse anti-PAI1 antibody (HA601152) at 1/1,000 dilution.

Lane 1: no heat NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: no heat NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate (20 µg/Lane)
Lane 5: HUVEC cell lysate (20 µg/Lane)
Lane 6: Mouse placenta tissue lysate (40 µg/Lane)
Lane 7: Rat placenta tissue lysate (40 µg/Lane)

Notice: no heat means the lysate is not boiled.

Predicted band size: 45 kDa
Observed band size: 45/100 kDa

Exposure time: 10 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601152) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (NBI02H) at 1:100,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Mouse anti-PAI1 antibody (HA601152) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601152) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human placenta tissue with Mouse anti-PAI1 antibody (HA601152) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601152) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunofluorescence analysis of paraffin-embedded human placenta tissue labeling PAI1 with Mouse anti-PAI1 antibody (HA601152) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA601152, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Flow cytometric analysis of HepG2 cells labeling PAI1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601152, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Flow cytometric analysis of NIH/3T3 cells labeling PAI1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601152, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).