Description:

Beta-Actin is highly conserved proteins and is ubiquitously expressed in all eukaryotic cells at a constant level regardless of experimental treatment or technical procedure in most cases. Therefore, measurement of beta-Actin is generally used as an internal control for experimental error. Signosis has provided b-actin sandwich ELISA assay specifically to detect the endogenous levels of beta-actin, which can be used as internal control for most of ELISA assays with human, mouse and rat samples.

Principle:

Beta-actin ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes a rabbit polyclonal beta-actin capture antibody for immobilization on the microtiter wells, and a mouse monoclonal beta-actin detection antibody along with anti-mouse conjugated to horseradish peroxidase (HRP) for detection. The test sample is allowed to react simultaneously with the two antibodies, resulting in the beta-actin molecules being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove unbound-labeled antibodies. A HRP substrate, TMB, is added to result in the development of a blue color. The color development is then stopped with the addition of stop solution changing the color to yellow. The concentration of beta-actin is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

 

 

 

Data:

Figure 1. Detection of beta-actin protein with beta-actin ELISA kit. HeLa cells were grown in a 12-well plate. Cell lysate was prepared with Cell Lysis Buffer, 1:2 serially diluted with 1 x Diluent buffer and subject to ELISA assay.

Signosis基因功能分析，靶标发现和确认，分析开发以及化合物筛选通常需要基于细胞的分析。经过工程改造以通过转基因整合到宿主基因组中表达目的基因的稳定细胞系提供了进行此类分析的有效方法。然而，由于建立这种细胞系的过程繁琐，开发过程耗时，劳动，试剂和材料的昂贵以及单个克隆的转染后验证，是否值得仍然值得商de在单个实验室中开发这种稳定细胞系的方法。  为了促进研究，Signosis现在提供了一系列基因表达或荧光素酶报告基因就绪的稳定细胞系。 萤光素酶报道基因稳定细胞系EGFR稳定表达细胞系HER2（ERBB2）稳定表达的细胞系pLenti-MSCV-EGFR-GFP载体pCMV-EGFR载体pCMV-EGFR-IRES-GFP载体pMSCV-EGFR-GFP载体EGFR突变荧光素酶表达载体GAL4-UAS-luc报告基因稳定细胞系NR LBD驱动的GAL4 Reporter稳定细胞系亲代细胞系