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磷酸化条带差?那是没有遇见它,PhosphoBLOCKER!

发布者:艾美捷科技    发布时间:2021-11-11     
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* 文末有干货下载:《艾美捷-WB常见问题-50问.PDF》

 

我有一个做蛋白质磷酸化(信号通路)研究的朋友,他说:WB是条不归路,WB难,磷酸化蛋白的WB更难!他曾创造过一个实验室记录,做了200多次磷酸化蛋白,出结果的只有60几次,与理论一致的只有40次不到。他,曾崩溃过...

 

磷酸化蛋白

 

现实情况确实如此,内源性目的蛋白本身在细胞裂解物中表达丰度不高,而其磷酸化修饰的蛋白含量就更低了!即使用高浓度抗体,延长曝光时间,依旧很难获得理想的效果。大多数市售的WB封闭剂(如牛奶,血清,BSA)虽然可以满足阻断膜上的未反应位点,减少检测过程中非特异性抗体的数量,但它们并不是为了在印迹过程中保护磷蛋白抗原而设计的,本身含有磷酸化蛋白成分,因而并不能为磷酸化WB雪中送炭!

 

作为专业的生命科学医药原料供应商,艾美捷科技为您推荐:磷酸化抗体专用:封闭液与抗体稀释液(二合一),让悲剧不再发生!

 

CellBiolabs

产品PhosphoBLOCKER Blocking Reagent
PhosphoBlocker封闭液
货号CBL-AKR-103
规格1L
产品形式50g 干粉混合物
溶解配置50g干粉溶解到1L的PBST/TBST溶液中,配制5% PhosphoBLOCKER 溶液
封闭液用途使用5% PhosphoBLOCKER 溶液,封闭膜
稀释液用途5% PhosphoBLOCKER溶液,可用于稀释一抗和二抗
说明书下载点击下载
运输条件常温运输
保存方法PhosphoBLOCKER溶液仅4℃保存1周,不可长期保存
注意事项深色颗粒的存在是我们制造过程中的正常现象,不会对产品的性能产生不利影响。如果需要,可以使用标准实验室滤纸过滤PhosphoBLOCKER?溶液,去除颗粒

*** 该产品仅用于科研实验.

 

  • PhosphoBlocker封闭液特点:

1.与奶粉,血清,BSA相比,更卓越的封闭效果

2.同时,可作为抗体稀释液使用,无需单独购买

3.增强低水平的磷蛋白信号而不增加背景

4.在Western Blot期间保留磷蛋白抗原

5.易于使用的预混合干混合物


  • PhosphoBlocker效果对比:

 

A549细胞裂解液,检测磷酸化-p38蛋白.

Fig. A549细胞裂解液,检测磷酸化-p38蛋白.

 

  • Cell也在用的磷酸化抗体专用封闭液:

Orthofer, Michael, et al. "Identification of ALK in Thinness." Cell181.6 (2020): 1246-1262.

 

Cell也在用的磷酸化抗体专用封闭液

【摘要】尽管世界大部分地区都存在同样致肥的环境,但个体对体重增加的易感性存在相当大的差异。尽管许多研究都集中在确定肥胖的遗传易感性,但我们在一个独特的表型爱沙尼亚队列中对代谢健康的瘦个体(全人群 BMI 谱的最低第 6 个百分位数)进行了 GWAS。我们发现间变性淋巴瘤激酶 (ALK) 作为候选瘦基因。在果蝇中,RNAi 介导的 Alk 敲低导致甘油三酯水平降低。在小鼠中,Alk 的基因缺失导致瘦弱的动物对饮食和瘦素突变引起的肥胖具有明显的抵抗力。从机制上讲,我们发现下丘脑神经元中的 ALK 表达通过交感神经控制脂肪组织脂肪分解来控制能量消耗。我们的遗传和机制实验将 ALK 鉴定为瘦身基因,该基因与抵抗体重增加有关。

 

Cell也在用的磷酸化抗体专用封闭液

western-blotting.jpg

 

更多已发表文章:

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艾美捷-WB常见问题-50问



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