Anti-uPA Receptor/U-PAR antibody [EPR22173-257] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] - BSA and Azide free (AB240825)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab221680).

• Flow Cyt

Unknown

Flow Cytometry - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] - BSA and Azide free (AB240825)

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)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221680).

• IP

Unknown

Immunoprecipitation - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] - BSA and Azide free (AB240825)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221680).

All lanes:

Immunoprecipitation - Anti-uPA Receptor/U-PAR antibody [EPR22173-257] (<a href='/en-us/products/primary-antibodies/upa-receptor-u-par-antibody-epr22173-257-ab221680'>ab221680</a>)

Predicted band size: 37 kDa

false