GLIPR2 Antibody / GLI pathogenesis-related protein 2 (FY12956)

Immunohistochemical staining of GLIPR2 using anti-GLIPR2 antibody. GLIPR2 was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-GLIPR2 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Immunohistochemical staining of GLIPR2 using anti-GLIPR2 antibody. GLIPR2 was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-GLIPR2 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Immunohistochemical staining of GLIPR2 using anti-GLIPR2 antibody. GLIPR2 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-GLIPR2 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Immunohistochemical staining of GLIPR2 using anti-GLIPR2 antibody. GLIPR2 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-GLIPR2 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of GLIPR2 using anti-GLIPR2 antibody. Lane 1: human U251 whole cell lysates, Lane 2: human U20S whole cell lysates, Lane 3: human THP-1 whole cell lysates, Lane 4: human Hela whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat RH35 whole cell lysates, Lane 8: mouse brain tissue lysates, Lane 9: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GLIPR2 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. A specific band was detected for GLIPR2 at approximately 17 kDa. The expected molecular weight of GLIPR2 is ~17 kDa.

Flow Cytometry analysis of THP-1 cells using anti-GLIPR2 antibody. Overlay histogram showing THP-1 cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GLIPR2 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Immunoprecipitating GLIPR2 in U251 whole cell lysate. Western blot analysis of GLIPR2 using anti-GLIPR2 antibody. Lane 1: U251 whole cell lysates (30ug) Lane 2: Rabbit control IgG instead of anti-GLIPR2 antibody in U251 whole cell lysate. Lane 3: anti-GLIPR2 antibody (2ug) + U251 whole cell lysate (500ug) After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-GLIPR2 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for GLIPR2 at approximately 17 kDa. The expected molecular weight of GLIPR2 is at 17 kDa.