siRNA / RNAi /miRNA transfection Mouse Glomerular Mesangial cells

Get tips on using Fabp5 siRNA to perform siRNA / miRNA gene silencing Rat - PC12 Fabp5

Get tips on using Vps13a siRNA to perform siRNA / miRNA gene silencing Rat - PC12 chorein

Get tips on using SignalSilence® PTEN siRNA to perform siRNA / miRNA gene silencing Human - A2780 PTEN

Get tips on using Pre-designed and validated siRNA against gene IGFBP1 to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1)

Get tips on using PACE4 siRNA (h) to perform siRNA / miRNA gene silencing Human - DuCaP

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

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 ADAMTS13 siRNA to perform siRNA / miRNA gene silencing Human - HUVEC ADAMTS-13

Get tips on using siRNA SKIL to perform siRNA / miRNA gene silencing Human - LuCaP-77 SKIL