siRNA / miRNA gene silencing Human Primary Endometrial Stromal Cells

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

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.

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 Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Immortalized cell lines iPSC

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Primary human breast tumors-Mammospheres

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

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human osteoblasts - rheumatoid arthritis

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