IDH2 Antibody – Mouse Monoclonal

Figure 1. Western blot analysis of IDH2 using anti-IDH2 antibody (MBS1753970).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates
Lane 2: human Sw620 whole cell lysates
Lane 3: human MCF-7 whole cell lysates
Lane 4: human HEPG2 whole cell lysates
Lane 5: human Jurkat whole cell lysates
Lane 6: rat heart tissue lysates
Lane 7: rat liver tissue lysates
Lane 8: rat PC-12 whole cell lysates
Lane 9: mouse heart tissue lysates
Lane 10: mouse NIH/3T3 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-IDH2 antigen affinity purified monoclonal antibody (Catalog # MBS1753970) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for IDH2 at approximately 45KD. The expected band size for IDH2 is at 45KD.

Figure 2. IF analysis of IDH2 using anti- IDH2 antibody (MBS1753970).
IDH2 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- IDH2 Antibody (MBS1753970) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 3. Flow Cytometry analysis of SiHa cells using anti- IDH2 antibody (MBS1753970).
Overlay histogram showing SiHa cells stained with MBS1753970 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-IDH2 Antibody (MBS1753970, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.