MultiTox-Glo Multiplex Cytotoxicity Assay

产品编号：G9272 品牌：promega 原厂货号：G9272
产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞毒性检测试剂盒
应用分类：

包装：

2 X 50ml

运保温度:

-20℃，避光

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其它组分：

Assay Buffer【子货号：G146B，包装：4 x 25ml，，运保温度：-20℃，避光】

AAF-Glo™ Substrate【子货号：G607B，包装：2 x 10ml，，运保温度：-20℃，避光】

GF-AFC Substrate【子货号：G608B，包装：2 x 50μl，，运保温度：-20℃，避光】

描述：

说明

MultiTox-Glo Multiplex Cytotoxicity Assay (MultiTox-Glo 叠加细胞毒性检测试剂盒) 是一种依次检测荧光和发光的试剂盒，可检测细胞群中活细胞和死细胞的相对数量。该试剂盒依次检测两种蛋白酶的活性。活细胞蛋白酶的活性仅存在于完整的活细胞，是通过一种可穿透细胞膜的荧光肽类底物(GF-AFC) 来实现检测的。这种底物进入完整的活细胞后被活细胞蛋白酶剪切释放出AFC，产生的荧光信号与活细胞数成正比。当细胞膜完整性丧失，这种活细胞蛋白酶活性标志物泄漏到周围培养物中，随即失活。试剂盒提供的第二种底物是生物发光的肽类底物AAF- 氨基萤光素(AAF-aminoluciferin)，不能穿透细胞，用于检测死细胞蛋白酶活性，死细胞相关的蛋白酶会从已经丧失细胞膜完整性的细胞中漏出，并剪切上述底物，释放出的游离氨基萤光素产物通过试剂盒提供的Ultra-Glo® 重组萤光素酶产生的 “辉光型”发光来检测。

MultiTox-Glo Assay 以比率法将细胞活力与细胞毒性的测定反相联系起来，与现有测定细胞活力与细胞毒性的方法相关联。活细胞与死细胞的比率与细胞数目无关，因此可被用于归一化数据。该方法使细胞活力与细胞毒性的检测方法互相补充，减少了由于移液和细胞凝集所产生的检测错误，也可作为内参来验证由于受试化合物或培养基组分造成的化学干扰所引起的检测错误。

特点

• 可同时检测培养体系中活细胞与死细胞的数量：依次添加试剂，是均质化的“加样- 混合- 检测”模式。

• 自带内对照，可将数据进行归一化处理：活细胞数与死细胞数之间的比例与细胞总数无关，因此可被用于归一化数据。

• 迅速识别更多的假阳性与假阴性结果：彼此独立的细胞活力与细胞毒性检测使两者互为对照。如果受试化合物对其中的一种检测方法产生了干扰，则另一种检测方法可以起到内对照的作用。

• 提高数据质量：降低假阳性( 或假阴性) 的统计概率，并可以消除荧光干扰的问题。

原厂资料：

The MultiTox-Glo Multiplex Cytotoxicity Assay is a sequential-reagent-addition fluorescent and luminescent assay that measures the relative number of live and dead cells in cell populations. The MultiTox-Glo Assay sequentially measures two protease activities; one is a marker of viability, and the other is a marker of cytotoxicity. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant, peptide substrate (GF-AFC). This substrate enters intact cells, where it is cleaved by the live cell protease activity to release AFC and generate a fluorescent signal that is proportional to the number of viable cells. The live-cell protease becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium. A second, luminogenic cell-impermeant peptide substrate (AAF-aminoluciferin) is used to measure dead-cell protease activity, which is released from cells that have lost membrane integrity. The liberated aminoluciferin product is measured as "glow type" luminescence generated by Ultra-Glo™ Recombinant Luciferase provided in the assay reagent.

The MultiTox-Glo Assay gives ratiometric, inversely correlated measures of cell viability and cytotoxicity, which correlate with established methods for measuring viability and cytotoxicity. The ratio of viable cells to dead cells is independent of cell number and, therefore, can be used to normalize data. Having complementary cell viability and cytotoxicity measures reduces errors associated with pipetting and cell clumping, as well as serving as an internal control to allow identification of errors resulting from chemical interference from test compounds or media components.

Features - Benefits

Measure the Number of Live Cells and Dead Cells in Culture: Sequential-reagent-addition assay with a homogeneous "add-mix-measure" protocol.
Normalize Data with a Built-In Internal Control: The ratio of the number of live cells/number of dead cells is independent of cell number and can be used to normalize data. This normalization makes results more comparable well-to-well, plate-to-plate and day-to-day.
Immediately Identify More False-Positives and False-Negatives: Independent cell viability and cytotoxicity measurements serve as controls for each other. If test compounds interfere with one assay chemistry, the other serves as an internal control.
Improve your Data: Reduce statistical probability of false-positives (or false-negatives), and eliminate fluorescence interference issues by luminescence readout.

Applications

Determination of cell viability and cytotoxicity.
Data normalization.

Notes

Using 100μl/assay in a 96-well plate format, Cat.# G9270 is sufficient to perform 100 assays; Cat.# G9271, 500 assays; Cat.# G9272, 1,000 assays. Using 25μl per well in a 384-well plate format, Cat.# G9270 is sufficient to perform 400 assays; Cat.# G9271, 2,000 assays; Cat.# G9272, 4,000 assays.