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>10次
| 产品编号 | 产品名称 | 产品包装 | 产品价格 |
| AA329 | Phospho-Akt(Ser473)抗体(兔多抗) | >10次 | 683.00元 |
| 来源 | 用途 | 交叉反应性 | 抗体识别位点 | Akt分子量 |
| Rabbit | WB, IP, F | H, M, R, C, Hm | Akt C-terminal | ~60kD |
WB, Western blot; IP, Immunoprecipitation; F, Flow cytometry.
H,human;M,mouse;R,rat; C,chick;Hm,hamster.
本Phospho-Akt(Ser473)抗体(Phospho-Akt(Ser473)antibody)为进口分装,用人工合成的小鼠Akt Ser473附近的一段多肽进行适当修饰后免疫rabbit,然后用protein A和抗原多肽亲和柱经过两步纯化得到的高纯度抗体。
本Phospho-Akt(Ser473)抗体识别Ser473被磷酸化的Akt1。相应位点被磷酸化的Akt2和Akt3也可以被本抗体识别。未发现本抗体可以识别其它位点磷酸化的Akt或磷酸化的其它蛋白例如磷酸化PKC、p70S6kinase等。
Akt又称PKB或Rac,在细胞存活和凋亡中起重要作用。胰岛素等生长和存活因子都可以激活Akt信号途径。Akt的Ser473可以被PDK2磷酸化。PI3 Kinase-Akt信号途径是一条经典的信号途径,LY294002等PI3 kinase的抑制剂抑制PI3 kinase时,通常就会抑制Akt激活。
配套提供了Western一抗稀释液,可以用于Western检测时的一抗稀释。
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 | F |
| 1:1000 | 1:100 | 1:25 |
本抗体如果用于常规的Western检测,至少可以检测10次。
包装清单:
| 产品编号 | 产品名称 | 包装 |
| AA329-1 | Phospho-Akt(Ser473)抗体(兔多抗) | 10μl |
| AA329-2 | Western一抗稀释液 | 10ml |
| - | 说明书 | 1份 |
保存条件:
Phospho-Akt(Ser473)抗体-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:
对于本抗体,Western检测时一抗要4℃缓慢摇动过夜,如果仅短时间与一抗孵育检测效果较差。
在Western实验后,请注意回收稀释的抗体。回收的抗体在进行Western实验时至少可以重复使用10次。稀释后的抗体,包括已经使用过的稀释抗体,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10000g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。
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。
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文献和实验1. Mu Zhang, Chunjie Yang, Meng Zhu, Li Qian, Yan Luo, Huimin Cheng, Rong Geng, Xiaojun Xu, Cheng Qian, Yu Liu . Saturated fatty acids entrap PDX1 in stress granules and impede islet beta cell function Diabetologia. 2021 May;64(5):1144-1157. doi: 10.1007/s00125-021-05389-4. (IF 7.518)
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5. Feng Y, Zou Z, Gao L, Zhang X, Wang T, Sun H, Liu Y, Chen X. . Umbilical Cord Blood-Derived Stromal Cells Regulate Megakaryocytic Proliferation and Migration Through SDF-1/PECAM-1 Pathway. Cell Biochem Biophys. 2012 Sep;64(1):5-15. (IF 2.073)
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8. Luo L, Lu AM, Wang Y, Hong A, Chen Y, Hu J, Li X, Qin ZH. . Chronic resistance training activates autophagy and reduces apoptosis of muscle cells by modulating IGF-1 and its receptors, Akt/mTOR and Akt/FOXO3a signaling in aged rats. Exp Gerontol. 2013 Apr;48(4):427-36. (IF 3.376)
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出来的.像前面几位战友说的那样,可以考虑加大上样量 2. 是否WB的条件不好? 可以跑其他蛋白的WB看看,如果其他蛋白质都跑出来的很好的话, 可以考虑换个抗体试一试,,但是话说回来,cell signaling的p-AKT 很好的,我也用了好几年.ser473和T308都不错的. 3. 条带弱的话,可以用 can get sinal 去提高抗体的敏感度. BTW, 我自己跑p-akt的时候, 1抗是1:1500 2抗是 1:20000 good luck
米宝 这俩天做实验,极其郁闷。我做的总AKt和pAkt,大鼠脑组织,第一次跑总AKT的时候,我觉得可能是因为蛋白是提取的浓度比较好吧,出来了一条浓浓的条带,三组总AKt都出了,接下来做pAkt,就出了一个带,效果挺明显的。后来再做pAkt的时候,跑了4张膜,显影的时候,太干净了,上面什么都没有,甚至连个杂带,斑点 都没!实验是在实验员的领导下完成的,技术上基本没什么纰漏,查阅本版面有关这问题方面的介绍,有的解释说pakt降解了!一抗是用的Phospho-Akt
receptorr 我在做HaCaT细胞的磷酸化Akt(473)时,背景比较高,该怎么办?(即未处理组的Akt比较高) 请高手支招。 谢谢 magichunter 说得太粗略了。 receptorr 我做的细胞是HaCaT 看看加入药物后HaCaT的磷酸化Akt是否有增高, 所以希望没有加药的那一组HaCaT的磷酸化Akt水平低 但是实际做时,发现没有











