Anti-uPA Receptor/U-PAR antibody [EPR22173-257]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] (AB221680)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U937 (human histiocytic lymphoma cell line) cells, untreated or treated with TPA (200 nM, 72 hours), labeling uPA Receptor/U-PAR with ab221680 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membranous staining in U937 cells treated with TPA (200 nM, 72 hours).

The expression of uPA Receptor/U-PAR is induced in PMA-stimulated U937 cells (PMID : 24999729).

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] (AB221680)

Flow cytometric analysis of U937 (human histiocytic lymphoma cell line) cell line treated with 200 nM TPA for 72 hours (red) or Untreated control (green) labeling uPA Receptor/U-PAR with ab221680 at 1/500 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

The expression of uPA Receptor/U-PAR is induced in PMA-stimulated U937 cells (PMID : 24999729)

• IP

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Immunoprecipitation - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] (AB221680)

uPA Receptor/U-PAR was immunoprecipitated from 0.35 mg of U937 (human histiocytic lymphoma cell line) treated with 200 nM TPA for 72 hours whole cell lysate with ab221680 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221680 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : U937 treated with 200 nM TPA for 72 hours whole cell lysate 10 μg (Input).

Lane 2 : ab221680 IP in U937 treated with 200 nM TPA for 72 hours whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221680 in U937 treated with 200 nM TPA for 72 hours whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 30 seconds.

The expression of uPA Receptor/U-PAR is induced in PMA-stimulated U937 cells and the molecular weight is consistent with what has been described in the literature (PMID : 24999729).

All lanes:

Immunoprecipitation - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] (ab221680)

Predicted band size: 37 kDa

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