Anti-uPA Receptor/U-PAR antibody [EPR26916-73]

• IHC

Supplier Data

Immunohistochemistry - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling uPA Receptor/U-PAR with ab307895 at 1/100 followed by ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on the perifollicular zone of mouse spleen (PMID : 11156692). The section was incubated with ab307895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling uPA Receptor/U-PAR with ab307895 at 1/100 followed by ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on glomeruli and the endothelial cells of mouse kidney (PMID : 11156692). The section was incubated with ab307895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

Immunohistochemical analysis of paraffin-embedded mouse head and neck squamous cell carcinoma tissue labeling uPA Receptor/U-PAR with ab307895 at 1/100 followed by ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on the mouse head and neck squamous cell carcinoma. The section was incubated with ab307895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

uPA Receptor/U-PAR was immunoprecipitated from 0.35 mg bEnd.3 (mouse brain endothelial cell) whole cell lysate with ab307895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : bEnd.3 whole cell lysate Lane 2 : ab307895 IP in bEnd.3 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307895 in bEnd.3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (ab307895) at 1/30 dilution

All lanes:

bEnd.3 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 37 kDa,65 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (ab307895) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 40 µg

Lane 2:

Mouse kidney tissue lysate at 40 µg

Lane 3:

Mouse spleen tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 37 kDa,65 kDa

true

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The molecular weight observed is consistent with what has been described in the literature (PMID : 19667118). Observed MW(kDa) 24 (deglycosylated form), 37-65(glycosylated form). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane 1-3 : 6 seconds, Lane4 : 37 seconds.

All lanes:

Western blot - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (ab307895) at 1/1000 dilution

Lane 1:

Untreated bEnd.3 whole cell lysate at 30 µg

Lane 2:

bEnd.3 whole cell lysate treated with Protein Deglycosylation MIX II at 30 µg

Lane 3:

RAW264.7 whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 24 kDa,37 kDa,65 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (AB307895)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression tissue : kidney and spleen.

All lanes:

Western blot - Anti-uPA Receptor/U-PAR antibody [EPR26916-73] (ab307895) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 37 kDa

Observed band size: 37 kDa,65 kDa

false

Exposure time: 180s