Site Directed Mutagenesis (SDM) Human Point mutation SH-SY5Y

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Primary cells CD4+ T cells

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using CD163 Antibody, anti-human, PE-Vio® 770, REAfinity™ to perform Flow cytometry Anti-bodies Human - CD163

Get tips on using Alexa Fluor® 488 anti-human CD127 (IL-7Rα) Antibody to perform Flow cytometry Anti-bodies Human - CD127

Get tips on using Alexa Fluor® 488 anti-human CD15 (SSEA-1) Antibody to perform Flow cytometry Anti-bodies Human - CD15