WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing.
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 | ChIP | ChIP-seq |
| 1:500-1:2000 | - | 1:50-1:200 | - | - | - | - | - |
抗体详细信息如下:
| About this Antibody | |
| Name | Phospho-mTOR (Ser2448) Rabbit Polyclonal Antibody |
| Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
| Isotype | IgG |
| Purification method | Affinity purification |
| Positive samples | NIH/3T3, C6 |
| Cellular location | Cytoplasm, Cytoplasmic side, Endoplasmic reticulum membrane, Golgi apparatus membrane, Lysosome, Mitochondrion outer membrane, Nucleus, PML body, Peripheral membrane protein |
| Customer validation | WB (African green monkey, Human, Rat) |
| About the Immunogen | |
| Immunogen | A synthetic phosphorylated peptide around S2448 of human mTOR (NP_004949.1). |
| Sequence | TDSYS |
| Gene ID | 2475 |
| Swiss Prot | P42345 |
| Synonyms | MTOR; FRAP; FRAP1; FRAP2; RAFT1; RAPT1; SKS; mechanistic target of rapamycin |
| Category | AMPK Signaling; Angiogenesis; Autophagy Signaling; B Cell Receptor Signaling |
| Background | The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth. When sufficient nutrients are available, mTOR responds to a phosphatidic acid-mediated signal to transmit a positive signal to p70 S6 kinase and participate in the inactivation of the eIF4E inhibitor, 4E-BP1. These events result in the translation of specific mRNA subpopulations. mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481. mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors. For these reasons, mTOR is currently under investigation as a potential target for anti-cancer therapy. |
包装清单：
| 产品编号 | 产品名称 | 包装 |
| XY-5869 | Phospho-mTOR (Ser2448) Rabbit Polyclonal Antibody | 50μl |
| XY-050 | Western一抗稀释液 | 50ml |
| — | 说明书 | 1份 |
保存条件：
Phospho-mTOR (Ser2448) Rabbit Polyclonal Antibody -20ºC保存，Western一抗稀释液-20ºC或4ºC保存，一年有效。Western一抗稀释液优先推荐4ºC保存，长期不使用可以考虑-20ºC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。
本产品于专业人员的科学研究用，不得用于临床诊断或ZL，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信裕生物提供的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。
2. 免疫染色：
可以使用信裕生物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，
3. 其它实验操作请自行参考适当的protocol进行。
4. 代表性图片：
 |  |
| Fig. 1. Western blot analysis of extracts of NIH/3T3 and C6 cells, using Phospho-MTOR-S2448 antibody at 1:2000 dilution. NIH/3T3 cells were treated by Insulin for 10 minutes after serum-starvation overnight. C6 cells were treated by IGF-1 for 30 minutes after serum-starvation overnight. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s. |
