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※サムネイル画像をクリックすると拡大画像が表示されます。
Figure 1. Western blot analysis of EIF5 using anti-EIF5 antibody (M04623).
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 K562 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human RT4 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-EIF5 antigen affinity purified monoclonal antibody (Catalog # M04623) 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 EIF5 at approximately 49 kDa. The expected band size for EIF5 is at 49 kDa.
Figure 2. Flow Cytometry analysis of SiHa cells using anti-EIF5 antibody (M04623).
Overlay histogram showing SiHa cells stained with M04623 (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-EIF5 Antibody (M04623, 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.
Figure 3. IF analysis of EIF5 using anti-EIF5 antibody (M04623).
EIF5 was detected in an immunocytochemical section of PC-3 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-EIF5 Antibody (M04623) 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.
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Figure 1. Western blot analysis of EIF5 using anti-EIF5 antibody (M04623).
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 K562 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human RT4 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-EIF5 antigen affinity purified monoclonal antibody (Catalog # M04623) 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 EIF5 at approximately 49 kDa. The expected band size for EIF5 is at 49 kDa.
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| 別品名 |
Thioredoxin, mitochondrial; MTRX; Mt-Trx; Thioredoxin-2; TXN2; TRX2
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| 種由来 |
Human
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| 交差種 |
Human
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| 免疫動物 |
Mouse
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| クローン |
2F13C4
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| 抗体クラス |
IgG2b
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| 標識物 |
Cyanine 3
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| 精製度 |
Affinity Purified
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| GENE ID |
1983
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| Accession No.(Gene/Protein) |
P55010
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| Gene Symbol |
EIF5
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| 参考文献 |
1. Das, S., Ghosh, R., Maitra, U. Eukaryotic translation initiation factor 5 functions as a GTPase-activating protein. J. Biol. Chem. 276: 6720-6726, 2001. 2. Hartz, P. A. Personal Communication. Baltimore, Md. 5/5/2010. 3. Jennings, M. D., Pavitt, G. D. eIF5 has GDI activity necessary for translational control by eIF2 phosphorylation. Nature 465: 378-381, 2010. Note: Erratum: Nature 468: 122 only, 2010.
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| メーカー |
品番 |
包装 |
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BBT
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M04623-CY3
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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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