Anti-IDH1 Antibody (monoclonal, 16H7)

• Catalog Number: orb623769
• Category: Antibodies
• Description: Anti-IDH1 Antibody (monoclonal, 16H7). Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG1
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Liquid
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human IDH1, different from the related mouse and rat sequences by one amino acid.
• UniProt ID: O75874
• MW: 47 kDa
• Tested applications: FC, ICC, IF, WB
• Application notes: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Antibody Type: Primary Antibody
• Clone Number: 16H7
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Cytosolic NADP isocitrate dehydrogenase antibody;
• Note: For research use only

Flow Cytometry analysis of CACO-2 cells using anti-IDH1 antibody. Overlay histogram showing CACO-2 cells (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 mouse anti-IDH1 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of IDH1 using anti-IDH1 antibody. IDH1 was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-IDH1 Antibody overnight at 4℃. DyLight488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37℃. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of IDH1 using anti-IDH1 antibody. 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 50 ug of sample under reducing conditions. Lane 1: human HepG2 tissue lysates, Lane 2: human Caco-2 whole cell lysates, Lane 3: human U-87MG whole cell lysates, Lane 4: human THP-1 whole cell lysates, Lane 5: human Hela whole cell lysates. Lane 6: human K562 whole cell lysates. Lane 7: human PC-3 whole cell lysates. Lane 8: human HEK293 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 mouse anti-IDH1antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4℃, 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 with Tanon 5200 system. A specific band was detected for IDH1 at approximately 47 KD. The expected band size for IDH1 is at 47 KD.

Western blot analysis of IDH1 using anti-IDH1 antibody. 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 50 ug of sample under reducing conditions. Lane 1: rat liver tissue lysates, Lane 2: rat RH35 whole cell lysates, Lane 3: mouse liver 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 mouse anti-IDH1antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4℃, 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 with Tanon 5200 system. A specific band was detected for IDH1 at approximately 47 KD. The expected band size for IDH1 is at 47 KD.