Anti-NRP2 antibody [EPR23808-72]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NRP2 antibody [EPR23808-72] (AB273584)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-87 MG cells labelling NRP2 with ab273584 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and membranous staining in U-87 MG cell line. Negative control : MCF7 (PMID : 18818766). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/202 dilution (Red). The Nuclear counterstain was DAPI (Blue).

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

• Flow Cyt

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Flow Cytometry - Anti-NRP2 antibody [EPR23808-72] (AB273584)

Flow cytometric analysis of MCF7 (Human breast adenocarcinoma epithelial cell, Left) / U-87 MG (Human glioblastoma-astrocytoma epithelial cell, Right) cells labelling NRP2 with ab273584 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : MCF7.

Gated on viable cells.

• IP

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Immunoprecipitation - Anti-NRP2 antibody [EPR23808-72] (AB273584)

NRP2 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate with ab273584 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273584 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate 10 ug

Lane 2 : ab273584 IP in U-87 MG whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab273584 in U-87 MG whole cell lysate

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

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-NRP2 antibody [EPR23808-72] (ab273584)

Predicted band size: 105 kDa

Observed band size: 130 kDa

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• WB

Lab

Western blot - Anti-NRP2 antibody [EPR23808-72] (AB273584)

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

The expression profile observed is consistent with what has been described in the literature (PMID : 18394556, 26156437).

Negative control : MCF 7 (PMID : 17912247).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-NRP2 antibody [EPR23808-72] (ab273584) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 40 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 105 kDa

Observed band size: 130 kDa

false