ELISA-based Ki-ras gene mutation analyses in pancreatic and cholangiocarcinoma cells and tissues.
Abstract
Detection of Ki-ras mutations is one possible modality for diagnosis of human neoplasms, particularly of the gastrointestinal tract. Little is known about the protein expression levels of ras p21. Here we show a semi-quantitative analysis of mutated ras p21 in protein extracts from pancreatic and cholangiocarcinoma cell lines and from tumor tissues. Comparison with DNA sequencing data confirmed the specificity of the ELISA-mediated mutation analysis. Epithelial cell content from stroma rich tissues was estimated by measuring the CK 8, 18 and 19 concentration in protein extracts from tissue samples comparing this with the average keratin content in extracts from 21 GI-carcinoma cell lines. The combination of both tests allowed the analysis of the ras p21 content of tissues with as little as 5% tumor cell content.
呵呵,这个ELISA检测基因突变比较意思。刚刚看了几篇文章,跟楼主分享一下。
文章:Detection of four b-thalassemia point mutations in Iranians using a PCR-ELISA genotyping system.
检测图中四个位置的基因突变。这个检测方法的四要素:a,DIG labed PCR products b,5’biotinylated oligos (probe) c,streptavidin-coated microtiter d,HRP conjugate anti-digoxigenin antibody 。 检测方法的本质是基于:a,allele-specific oligonucleotide hybridization b,biotin-streptavidin system c,Enzyme-linked immunosorbent assay
Fig 1 - Detection of four b-thalassemia point mutations in Iranians using a PCR-ELISA genotyping system.
文章Molecular Detection of Galactosemia Mutations by PCR-ELISA 有step by step的protocol。
Fig 2 - Molecular Detection of Galactosemia Mutations by PCR-ELISA
另外基于相同原理的技术还有PPT-ELISA。这是用来筛截短突变的,相关文章见:1,Rapid screen for truncating ATM mutations by PTT-ELISA . 2,An ELISA-based high throughput protein truncation test for inherited breast cancer .
Fig 3 - Rapid screen for truncating ATM mutations by PTT-ELISA
Fig 4 - An ELISA-based high throughput protein truncation test for inherited breast cancer
Fig2 - Molecular Detection of Galactosemia Mutations by PCR-ELISA
Fig3 - Rapid screen for truncating ATM mutations by PTT-ELISA
Fig4 - An ELISA-based high throughput protein truncation test for inherited breast cancer