PathScan? Total Cyclin D1 Sandwich ELISA Kit

• 产品编号：CST-7918C      品牌：CST       原厂货号：7918C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

PathScan® Total Cyclin D1 Sandwich ELISA Kit能够检测人、猴、小鼠和大鼠细胞中内源性水平的细胞周期蛋白D1，见图1.试剂盒的灵敏性见图2. 本试剂盒用来检测指定物种的蛋白，即通过内部测试确定的指定物种蛋白，也可能检测来自其他物种的同源蛋白。PathScan® Total Cyclin D1 Sandwich ELISA Kit是固相夹心酶联免疫吸附试验（ELISA），能够检测内源性水平的总细胞周期蛋白D1。细胞周期蛋白D1兔抗包被微孔板。细胞裂解液孵育后，磷酸和非磷酸化的细胞周期蛋白D1都能被包被的抗体捕获。充分冲洗后，加入细胞周期蛋白D1小鼠检测抗体检测捕获的酪氨酸磷酸化的细胞周期蛋白D1。随后用抗鼠IgG，HRP-标记的抗体识别结合的检测蛋白。加入HRP底物TMB进行显色。显色的吸光度值与总的细胞周期蛋白D1的量成正比。试剂盒中的抗体是针对试剂盒定制的。细胞周期蛋白依赖性激酶CDK4和CDK6的活性受T-环磷酸化、许多细胞周期蛋白伴侣（D型细胞周期蛋白）和与CDK抑制剂Cip/Kip or INK 蛋白家族的联合调节（1）。细胞周期蛋白D/CDK4和p27 Kip1无活性三元复合物需要胞外促有丝分裂信号刺激，从而释放释放和降解p27，随后细胞周期蛋白D水平上升，通过限制点和Rb-依赖性的进入S期来影响细胞进程（2）。细胞周期蛋白D/CDK4的活性复合物靶向视网膜母细胞瘤蛋白并将其磷酸化，使转录因子E2F释放，激活G1/S期基因的表达（3）。伴随着生长因子消失，通过下调蛋白表达和磷酸化依赖性的降解，下调细胞周期蛋白D的表达水平（4）。

1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
2. Sherr, C.J. (1996) Science 274, 1672-1677.
3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

原厂资料：

Specificity / Sensitivity

PathScan® Total Cyclin D1 Sandwich ELISA Kit detects endogenous levels of cyclin D1 protein in human, monkey, mouse, and rat cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Total Cyclin D1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total cyclin D1 protein. A Cyclin D1 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, both phospho and nonphospho cyclin D1 proteins are captured by the coated antibody. Following extensive washing, Cyclin D1 Mouse Detection Antibody is added to detect the captured cyclin D1 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of total cyclin D1 protein. 
 Antibodies in kit are custom formulations specific to kit.

Background

Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to affect progression through the restriction point and Rb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of protein expression and phosphorylation-dependent degradation (4).

1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
2. Sherr, C.J. (1996) Science 274, 1672-1677.
3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

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).