IDH3B Antibody / Isocitric dehydrogenase subunit beta (FY13009)

Immunohistochemical staining of IDH3B using anti-IDH3B antibody. IDH3B was detected in a paraffin-embedded section of follicles of human thyroid 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-IDH3B 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 IDH3B using anti-IDH3B antibody. Lane 1: human LANAP whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human RT4 whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse brain tissue lysates, Lane 8: mouse heart tissue 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-IDH3B antibody at 0.25 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 IDH3B at approximately 42 kDa. The expected molecular weight of IDH3B is ~42 kDa.

Immunohistochemical staining of IDH3B using anti-IDH3B antibody. IDH3B was detected in a paraffin-embedded section of follicles of human thyroid 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-IDH3B 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 IDH3B using anti-IDH3B antibody. IDH3B was detected in a paraffin-embedded section of follicles of human thyroid 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-IDH3B 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 IDH3B using anti-IDH3B antibody. IDH3B was detected in a paraffin-embedded section of follicles of human thyroid 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-IDH3B 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.

Flow Cytometry analysis of JK cells using anti-IDH3B antibody. Overlay histogram showing JK 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-IDH3B 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 (Red line) was also used as a control.

Immunoprecipitating IDH3B in U251 whole cell lysate. Western blot analysis of IDH3B using anti-IDH3B antibody. Lane 1: U251 whole cell lysates (30ug) Lane 2: Rabbit control IgG instead of anti-IDH3B antibody in U251 whole cell lysate. Lane 3: anti-IDH3B antibody (2ug) + U251 whole cell lysate (500ug) After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-IDH3B 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 IDH3B at approximately 42 kDa. The expected molecular weight of IDH3B is at 42 kDa.