| 抗原部位 |
a.a.26-907
|
| 種由来 |
Human
|
| 標識物 |
iFluorTM 647
|
| 精製度 |
Affinity Purified
|
| 適用 |
Western Blot
Enzyme Linked Immunosorbent Assay
Immunohistochemistry
Flow Cytometry
|
| 免疫動物 |
Rabbit
|
| 交差種 |
Human
Mouse
Rat
|
| GENE ID |
8549
|
| Accession No.(Gene/Protein) |
O75473
|
| Gene Symbol |
LGR5
|
| 形状 |
凍結乾燥品
|
| 参考文献 |
1. Barker, N., Ridgway, R. A., van Es, J. H., van de Wetering, M., Begthel, H., van den Born, M., Danenberg, E., Clarke, A. R., Sansom, O. J., Clevers, H. Crypt stem cells as the cells-of-origin of intestinal cancer. Nature 457: 608-611, 2009.
2. Barker, N., van Es, J. H., Kuipers, J., Kujala, P., van den Born, M., Cozijnsen, M., Haegebarth, A., Korving, J., Begthel, H., Peters, P. J., Clevers, H. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449: 1003-1007, 2007.
3. de Lau, W., Barker, N., Low, T. Y., Koo, B.-K., Li, V. S. W., Teunissen, H., Kujala, P., Haegebarth, A., Peters, P. J., van de Wetering, M., Stange, D. E, van Es, J. E., Guardavaccaro, D., Schasfoort, R. B. M., Mohri, Y., Nishimori, K., Mohammed, S., Heck, A. J. R., Clevers, H. Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling. Nature 476: 293-297, 2011.
|
|
※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 LGR5 using anti-LGR5 antibody (A00239-2).
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 30 ug of sample under reducing conditions.
Lane 1: human SH-SY5Y whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human RT4 whole cell lysates,
Lane 4: human 293T 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 rabbit anti-LGR5 antigen affinity purified polyclonal antibody (Catalog # A00239-2) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for LGR5 at approximately 130 kDa. The expected band size for LGR5 is at 100,97,92 kDa.
Figure 2. IHC analysis of LGR5 using anti-LGR5 antibody (A00239-2).
LGR5 was detected in a paraffin-embedded section of mouse small intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-LGR5 Antibody (A00239-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 3. IHC analysis of LGR5 using anti-LGR5 antibody (A00239-2).
LGR5 was detected in a paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-LGR5 Antibody (A00239-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 4. Flow Cytometry analysis of 293T cells using anti-LGR5 antibody (A00239-2).
Overlay histogram showing 293T cells stained with A00239-2 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-LGR5 Antibody (A00239-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLightR488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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 1. Western blot analysis of LGR5 using anti-LGR5 antibody (A00239-2).
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 30 ug of sample under reducing conditions.
Lane 1: human SH-SY5Y whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human RT4 whole cell lysates,
Lane 4: human 293T 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 rabbit anti-LGR5 antigen affinity purified polyclonal antibody (Catalog # A00239-2) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for LGR5 at approximately 130 kDa. The expected band size for LGR5 is at 100,97,92 kDa.
|
|
|
| メーカー |
品番 |
包装 |
|
BBT
|
A00239-2-IFLUOR647
|
100 UG
|
※表示価格について
| 当社在庫 |
なし
|
| 納期目安 |
1週間程度
|
| 保存温度 |
-２０℃
|
|
