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Figure 1. Western blot analysis of DHODH using anti-DHODH antibody (M04035). 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 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: rat liver tissue lysates, Lane 7: mouse testis tissue lysates, Lane 8: mouse liver 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 mouse anti-DHODH antigen affinity purified monoclonal antibody (Catalog # M04035) at 0.5 μg/mL overnight at 4°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 # EK1001) with Tanon 5200 system. A specific band was detected for DHODH at approximately 43 kDa. The expected band size for DHODH is at 43 kDa.
Figure 2. IF analysis of DHODH using anti-DHODH antibody (M04035). DHODH was detected in an immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-DHODH Antibody (M04035) overnight at 4°C. DyLightR488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°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 U937 cells using anti-DHODH antibody (M04035). Overlay histogram showing U937 cells stained with M04035 (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-DHODH Antibody (M04035, 1 μg/1x106 cells) for 30 min at 20°C. DyLightR488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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Figure 1. Western blot analysis of DHODH using anti-DHODH antibody (M04035). 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 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: rat liver tissue lysates, Lane 7: mouse testis tissue lysates, Lane 8: mouse liver 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 mouse anti-DHODH antigen affinity purified monoclonal antibody (Catalog # M04035) at 0.5 μg/mL overnight at 4°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 # EK1001) with Tanon 5200 system. A specific band was detected for DHODH at approximately 43 kDa. The expected band size for DHODH is at 43 kDa.
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| 別品名 |
SUMO-activating enzyme subunit 2; Anthracycline-associated resistance ARX; Ubiquitin-like 1-activating enzyme E1B; Ubiquitin-like modifier-activating enzyme 2; UBA2; SAE2, UBLE1B; HRIHFB2115
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| 種由来 |
Human
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| 交差種 |
Human Mouse Rat
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| 免疫動物 |
Mouse
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| クローン |
4E3
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| 抗体クラス |
IgG2b
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| 標識物 |
Allophycocyanin
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| 精製度 |
Affinity Purified
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| GENE ID |
1723
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| Accession No.(Gene/Protein) |
Q02127
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| Gene Symbol |
DHODH
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| 参考文献 |
1. Ng, S. B., Buckingham, K. J., Lee, C., Bigham, A. W., Tabor, H. K., Dent, K. M., Huff, C. D., Shannon, P. T., Jabs, E. W., Nickerson, D. A., Shendure, J., Bamshad, M. J. Exome sequencing identifies the cause of a mendelian disorder. Nature Genet. 42: 30-35, 2010. 2. White, R. M., Cech, J., Ratanasirintrawoot, S., Lin, C. Y., Rahl, P. B., Burke, C. J., Langdon, E., Tomlinson, M. L., Mosher, J., Kaufman, C., Chen, F., Long, H. K., Kramer, M., Datta, S., Neuberg, D., Granter, S., Young, R. A., Morrison, S., Wheeler, G. N., Zon, L. I. DHODHmodulates transcriptional elongation in the neural crest and melanoma. Nature 471: 518-522, 2011.
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| メーカー |
品番 |
包装 |
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BBT
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M04035-APC
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100 UG
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※表示価格について
| 当社在庫 |
なし
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| 納期目安 |
1週間程度
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| 保存温度 |
-20℃
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