Oxidative Stress Induced Gene Expression High Throughput Evaluation Solutions
Cellular systems generate reactive oxygen species during metabolic processes. Oxidative stress leads to activation or repression of many genes in important signaling pathways. DNA microarrays allow for high throughput assessment of changes in gene expression levels in any biological system.
Creative Bioarray offers a solution to study transcriptional changes in redox-engineered cell lines using gene expression microarrays. This solution can be used to assess gene expression changes induced by oxidative stress in mammalian cells and tissue samples in high throughput and is highly applicable to other biological systems that can be used to assess oxidative stress.
High Throughput Evaluation of Oxidative Stress Induced Gene Expression
Step 1. Measurement of oxidant levels by flow cytometry
Oxidation rates of probes for various redox-engineered cell lines by FACS analysis of cells harvested after treatment with PBS-EDTA
Step 2. Cell culture and cell harvesting for RNA isolation.
Step 3. RNA isolation using Qiagen purification procedure
The specific RNA isolation you choose will depend on your downstream application. Typically, any of the microarray, RT-PCR or Northern blotting methods are acceptable. Qiagen centrifugal column purification offers the advantage of performing an optional DNaseI digestion along with the purified RNA, thus avoiding further processing. However, detection of RNA molecules of 200 bp or smaller will be limited if the Qiagen purification procedure is used, and is therefore not recommended if you intend to use the RNA for miRNA analysis.
Step 4. RNA quality assessment
The analysis can measure the molecular weight distribution of the isolated RNA. If the sample is to be used for downstream microarray or QPCR experiments, the RNA integrity number (RIN) score should be between 7 and 10.
Step 5. Cellular mRNA expression analysis
Step 6. Synthesis of labeled cRNA and microarray hybridization
Step 7. Analysis of gene expression data obtained using microarrays
Step 8. Quantitative PCR validation of microarray data
Why Choose Us
- We use cell culture grade media for all cell culture techniques in our service. Reagents for nucleic acid and protein biochemical manipulations are purchased as DNase-free, RNase-free, pyrogen-free.
- The solution can be used for high-throughput assessment of gene expression changes induced by oxidative stress in mammalian cells and tissue samples and is highly applicable to other biological systems that can be used to assess oxidative stress.
- One can also add different concentrations of oxidants such as H2O2 or oxidant-generated compounds such as N,N'-dimethyl-4,4'-dichloropyridine (paraquat) for superoxide or diethylenetriamine NONOate (Deta NONOate) for NO in different cellular systems.
Creative Bioarray is dedicated to providing high-quality products, comprehensive services, and tailored solutions to support and facilitate life sciences and pharmaceutical research and development. If you have any questions or needs, please contact us, and our customer service staff will help you the first time.
All services and products are for scientific use only, not for medical use！
