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Figure 1. Western blot analysis of RPA70 using anti-RPA70 antibody (M01317-2).
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: HELA whole cell lysates,
Lane 2: MCF-7 whole cell lysates,
Lane 3: K562 whole cell lysates,
Lane 4: COS-7 whole cell lysates,
Lane 5: Caco-2 whole cell lysates,
Lane 6: A549 whole cell lysates,
Lane 7: HEPG2 whole cell lysates,
Lane 8: PC-3 whole cell lysates,
Lane 9: HEPA1-6 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-RPA70 antigen affinity purified monoclonal antibody (Catalog # M01317-2) 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 1:5000 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 RPA70 at approximately 12KD. The expected band size for RPA70 is at 12KD.
Figure 2. IHC analysis of RPA70 using anti-RPA70 antibody (M01317-2).
RPA70 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-RPA70 Antibody (M01317-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 3. IHC analysis of RPA70 using anti-RPA70 antibody (M01317-2).
RPA70 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-RPA70 Antibody (M01317-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 4. IHC analysis of RPA70 using anti-RPA70 antibody (M01317-2).
RPA70 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-RPA70 Antibody (M01317-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 5. IF analysis of RPA70 using anti-RPA70 antibody (M01317-2).
RPA70 was detected in immunocytochemical section of A549 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 2μg/mL mouse anti-RPA70 Antibody (M01317-2) overnight at 4°C. DyLight488 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.
6. Flow Cytometry analysis of A431 cells using anti-RPA70 antibody (M01317-2).
Overlay histogram showing A431 cells stained with M01317-2 (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-RPA70 Antibody (M01317-2,1μg/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°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.
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Figure 1. Western blot analysis of RPA70 using anti-RPA70 antibody (M01317-2).
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: HELA whole cell lysates,
Lane 2: MCF-7 whole cell lysates,
Lane 3: K562 whole cell lysates,
Lane 4: COS-7 whole cell lysates,
Lane 5: Caco-2 whole cell lysates,
Lane 6: A549 whole cell lysates,
Lane 7: HEPG2 whole cell lysates,
Lane 8: PC-3 whole cell lysates,
Lane 9: HEPA1-6 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-RPA70 antigen affinity purified monoclonal antibody (Catalog # M01317-2) 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 1:5000 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 RPA70 at approximately 12KD. The expected band size for RPA70 is at 12KD.
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| 別品名 |
Replication protein A 70 kDa DNA-binding subunit; RP-A p70; Replication factor A protein 1; RF-A protein 1; Single-stranded DNA-binding protein; Replication protein A 70 kDa DNA-binding subunit, N-terminally processed; RPA1; REPA1; RPA70
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| 種由来 |
Human
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| 交差種 |
Human
Mouse
Monkey
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| 適用 |
Western Blot
Immunohistochemistry
Immuno Fluorescence
Flow Cytometry
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| 免疫動物 |
Mouse
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| クローン |
11H4
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| 抗体クラス |
IgG2b
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| 抗原部位 |
C-terminus
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| 精製度 |
Affinity Purified
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| GENE ID |
6117
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| Accession No.(Gene/Protein) |
P27694
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| Gene Symbol |
RPA1
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| 分子量 |
12 kDa
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| 概要 |
Boster Bio Anti-RPA70 RPA1 Antibody Picoband® (monoclonal, 11H4) catalog # M01317-2. Tested in Flow Cytometry, IF, IHC, WB applications. This antibody reacts with Human, Monkey, Mouse. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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| 参考文献 |
1. Erdile, L. F., Heyer, W.-D., Kolodner, R., Kelly, T. J. Characterization of a cDNA encoding the 70-kDa single-stranded DNA-binding subunit of human replication protein A and the role of the protein in DNA replication. J. Biol. Chem. 266: 12090-12098, 1991. Note: Erratum: J. Biol. Chem. 268: 2268 only, 1993. 2. Haring, S. J., Mason, A. C., Binz, S. K., Wold, M. S. Cellular functions of human RPA1: multiple roles of domains in replication, repair, and checkpoints. J. Biol. Chem. 283: 19095-19111, 2008.
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| メーカー |
品番 |
包装 |
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BBT
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M01317-2
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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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| 法規制 |
毒
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| 保存温度 |
-２０℃
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