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| 产品编号 | 产品名称 | 产品包装 | 产品价格 |
| AF2053 | Topoisomerase I Rabbit Monoclonal Antibody | 50μl | 709.00元 |
| 来源 | 用途 | 交叉反应性 | 分子量 |
| Rabbit | WB, IF, IHC, ICC, FC | H, M | 91.0kDa |
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC,
Immunocytochemistry; FC, Flow Cytometry; ELISA, Enzyme-linked Immunosorbent Assay; ChIP, Chromatin
Immunoprecipitation Assay.
H, Human; M, Mouse; R, Rat; C, Chicken; Cw, Cow; Dg, Dog; Gp, Guinea pig; Hm, Hamster; Hr, Horse; Mk,
Monkey; Pg, Pig; Rb, Rabbit; S, Sheep; Z, Zebrafish; All, all species expected.
配套提供了Western一抗稀释液，可以用于Western检测或其它适当用途时的一抗稀释。
建议抗体使用时的稀释比例如下(实际使用时需根据抗原水平的高低作适当调整)：
| WB | IP | IF | IHC | ICC | FC | ELISA | ChIP |
| 1:1,000-1:5,000 | - | 1:50-1:500 | 1:50-1:200 | 1:50-1:500 | 1:50-1:100 | - | - |
抗体详细信息如下:
| About this Antibody | |
| Name | Topoisomerase I Rabbit Monoclonal Antibody |
| Category | Rabbit Monoclonal Antibody (RabMAb); Primary antibody |
| Isotype | IgG |
| Purification | Affinity purification |
| About the Immunogen | |
| Immunogen | Recombinant protein |
| Gene ID | 7150(Human); 21969(Mouse) |
| SwissProt | P11387(Human); Q04750(Mouse) |
| Synonyms | TOPI |
| Category | DNA Damage |
| Background | DNA topoisomerases play essential roles in many DNA metabolic processes including DNA repair. Topoisomerases can introduce DNA damage upon exposure to drugs that stabilize the covalent protein-DNA intermediate of the topoisomerase reaction. Lesions in DNA are also able to trap topoisomerase-DNA intermediates. DNA topoisomerase I (Top1) catalyzes the relaxation of supercoiled DNA by a mechanism of transient DNA strand cleavage characterized by the formation of a phosphotyrosyl bond between the DNA end and active site tyrosine. The antitumor agent camptothecin (CPT) reversibly stabilizes the covalent enzyme-DNA intermediate by inhibiting DNA religation. When a replication fork collides with a DNA Top1 cleavage complex, the covalently bound enzyme must be removed from the DNA 3' end before recombination-dependent replication restart. The tyrosyl-DNA phosphodiesterase Tdp1 and the structure-specific endonuclease Rad1-Rad10 function as primary alternative pathways of Top1 repair in Saccharomyces cerevisiae. In the budding yeast S. cerevisiae, DNA topoisomerases I and II can functionally substitute for each other in removing positive and negative DNA supercoils. |
包装清单:
| 产品编号 | 产品名称 | 包装 |
| AF2053 | Topoisomerase I Rabbit Monoclonal Antibody | 50μl |
| AZ050 | Western一抗稀释液 | 50ml |
| - | 说明书 | 1份 |
保存条件:
Topoisomerase I Rabbit Monoclonal Antibody -20℃保存，Western一抗稀释液-20℃或4℃保存，一年有效。Western一抗稀释液优先推荐4℃保存，长期不使用可以考虑-20℃保存，但冻融可能会导致出现轻微的浑浊和少量不溶物。
注意事项:
如果本抗体用于Western blot (WB)、免疫荧光(IF)、免疫细胞化学(ICC)等实验，请注意回收使用过的稀释抗体。回收的抗体通常至少可以重复使用5-10次。稀释后的抗体，包括已经使用过的稀释抗体，请4℃保存。
回收后重复使用的抗体，使用方法同新鲜稀释的抗体。如果在重复使用过程中发现抗体出现轻微混浊现象，可以10,000g离心1-3分钟，取上清用于后续检测。如果回收的抗体出现明显的絮状物或长霉长菌等情况，则可以考虑废弃该抗体。
提供的Western一抗稀释液也可以用于免疫荧光(IF)、免疫组化(IHC)、免疫细胞化学(ICC)等适当用途。如果希望获得最佳的检测效果，请考虑使用上述检测专用的一抗稀释液。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
使用说明:
请根据抗体的实际用途选择相应的使用方法。
1. Western检测：
a. 按照推荐的稀释比例用碧云天提供的Western一抗稀释液稀释抗体。
b. 把经过封闭的蛋白膜与稀释好的一抗4℃缓慢摇动过夜或室温缓慢摇动2小时，确保稀释的抗体至少能在摇动的瞬间覆盖蛋白膜。
c. 回收稀释的一抗，4℃保存以备下次继续使用。
d. 按照Western的实验步骤进行后续的洗涤、二抗孵育、洗涤和检测等操作。具体操作可以参考如下网页： http://www.beyotime.com/support/western.htm
2. 免疫染色：
可以使用碧云天生产的免疫染色一抗稀释液(P0103)稀释抗体，使用后注意回收稀释好的一抗，具体操作可以参考如下网页： http://www.beyotime.com/support/immunol-staining.htm
3. 其它实验操作请自行参考适当的protocol进行。
4. 代表性图片：
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| Fig. 1. Western blot analysis of TOP1 on different lysates using anti-TOP1 antibody at 1/1,000 dilution. Positive control: Lane 1: HepG2; Lane 2: Jurkat; Lane 3: MCF-7 | Fig. 2. ICC staining TOP1 in SHG-44 cells (green). The nuclear counter stain is DAPI (blue).Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X-100/PBS. | |
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| Fig. 3. Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-TOP1 antibody. Counter stained with hematoxylin. | Fig. 4. Flow cytometric analysis of HepG2 cells with TOP1 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody. |
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