Description:

Angiogenesis provides the crucial link between the avascular and vascular states and is a key event for sustained tumor growth and cancer progression. Angiogenesis as a biological switch process is governed by numerous pro- and anti-angiogenic factors, including vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGFb), epidermal growth factor (EGF), and transforming growth factor-beta (TGF-b). The mechanism of action of each of these factors is different, as are their origin and the stimuli for their production. The angiogenic switch refers to the balance between pro- and anti- angiogenic factors. Therefore, profiling of these factors is critical to understanding angiogenesis. Signosis’ Angiogenesis ELISA Strip I Profiling Assay allows simultaneously profiling 8 angiogenesis cytokines. Each well of the strip is coated with a primary antibody against a specific angiogenesis protein and total 8 wells of a strip target 8 different proteins. The difference of these proteins between two samples can be determined through data comparison.

Principle:

Each well of the strip is coated with a specific capture antibody to detect its corresponding cytokine in the sample. Therefore, 8 different proteins can be measures simultaneously. The test sample reacts simultaneously with two antibodies, resulting in the cytokines being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove unbound-labeled antibodies. The HRP substrate, TMB, is then added and causes a blue color change. The reaction is then terminated with Stop Solution, resulting in a yellow color. The concentrations of oxidative stress cytokines are directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm. The expression levels of these cytokines can be quantitatively compared between samples.

 

 

 

Data:

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Human Angiogenesis ELISA strip I for profiling 8 cytokines. HeLa cells were treated with (red bars) or without (light blue) 10ng/ml TNF. Conditioned media were collected (top) and cell lysates were prepared (bottom) for ELISA strip profiling.

 

 

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