PathScan? Acetyl-α-Tubulin Sandwich ELISA Kit

• 产品编号：CST-7204C      品牌：CST       原厂货号：7204C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

CST's PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit #7204检测内源性乙酰化α-Tubulin蛋白。如图1所示，使用Acetyl-α-Tubulin Sandwich ELISA Kit #7204，当使用TSA处理后，在COS细胞中一个高水平的乙酰化α-tubulin蛋白被检测。通过免疫印迹（western blot）分析总蛋白(乙酰化的和非乙酰化的)的水平仍然没有改变(图1)。当NIH/3T3和Jurkat细胞使用TSA处理后，类似的结果被获得(数据未显示)。因为通过内部实验确定，该试剂盒检测已明确的物种，但是可能也会检测其它物种的同源性蛋白。CST's PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit是一个固相夹心酶联免疫吸附法，其能检测内源性乙酰化的α-tubulin蛋白。一个α-tubulin mouse mAb已经被包被在微孔板上。在与细胞裂解液一起孵化后，α-tubulin蛋白(乙酰化的和非乙酰化的)被包被的抗体吸附。在大范围清洗之后，一个acetyl-lysine rabbit Ab被加入去检测已被吸附的乙酰化的α-tubulin蛋白。然后，Anti-rabbit IgG, HRP-linked Antibody #7074被用于识别已结合的检测抗体。HRP底物（TMB）加入后显色。该显色的吸光度值的大小与在乙酰化的α-tubulin蛋白量成正比例。试剂盒里的抗体是针对该试剂盒特定制备的。细胞骨架由三种类型的细胞质基质纤维组成：微管、微丝(肌动蛋白纤维)和中间纤维。球形微管蛋白亚基包含微管建筑块，由α/β-tubulin异源二聚体形成所有真核细胞的tubulin亚单位。γ-tubulin被需要在tubulin亚单位的成核聚合作用中去形成微管聚合物。许多细胞运动是通过微管动力介导的，包括纤毛和鞭毛的跳动、膜泡的核质转运、在减数分裂/有丝分裂期间染色体排布和神经轴突的迁移。这些运动由于竞争性微管聚合作用和解聚作用或通过微管动力蛋白的运动(1)。

1. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit #7204 detects endogenous levels of acetylated α-tubulin. As shown in Figure 1 using the Acetyl-α-Tubulin Sandwich ELISA Kit #7204, a high level of acetylated α-tubulin is detected in COS cells when treated with TSA. The level of total α-tubulin (acetylated and non-acetylated) remains unchanged as shown by Western analysis (Figure 1). Similar results are obtained when NIH/3T3 and Jurkat cells are treated with TSA (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated α-tubulin protein. An α-tubulin mouse mAb has been coated onto the microwells. After incubation with cell lysates, α-tubulin protein (acetylated and non-acetylated) is captured by the coated antibody. Following extensive washing, an acetyl-lysine rabbit Ab is added to detect the acetylated α-tubulin protein. Anti-rabbit IgG, HRP-linked Antibody #7074 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of acetylated α-tubulin protein.

Background

The cytoskeleton consists of three types of cytosolic fibers: microtubules, microfilaments (actin filaments), and intermediate filaments. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. γ-tubulin is required to nucleate polymerization of tubulin subunits to form microtubule polymers. Many cell movements are mediated by microtubule action, including the beating of cilia and flagella, cytoplasmic transport of membrane vesicles, chromosome alignment during meiosis/mitosis, and nerve-cell axon migration. These movements result from competitive microtubule polymerization and depolymerization or through the actions of microtubule motor proteins (1).

1. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).