siRNA / RNAi /miRNA transfection Human Cells Primary splenocytes

Get tips on using FlexiTube GeneSolution GS7052 for TGM2 to perform siRNA / miRNA gene silencing Human - Caki-2 TGM2

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 RNAqueous™ Total RNA Isolation Kit to perform RNA isolation / purification Cells - primary human lung fibroblasts

Get tips on using Orai1 Rat siRNA Oligo Duplex (Locus ID 304496) to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Orai1

Get tips on using ON-TARGET plus Mouse Becn1 (56208) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 4T1 BECN-1

Get tips on using ZR RNA MiniPrepTM kit to perform RNA isolation / purification Cells - primary human islets of langerhans

Get tips on using High Pure RNA Isolation Kit to perform RNA isolation / purification Cells - primary human dermal fibroblasts

Get tips on using AllPrep DNA/RNA Mini Kit to perform RNA isolation / purification Cells - primary human CD14+ monocytes

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human peripheral blood monocytes