siRNA / RNAi /miRNA transfection Human Cells A549

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Primary cells Human aortic smooth muscle cells (HOSMC)

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 Silencer® Select_Vamp2 siRNA(r) to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Vamp2

Get tips on using Silencer® Select_Vamp7 siRNA(r) to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Vamp7

Get tips on using IGF-IRα/β siRNA (r) to perform siRNA / miRNA gene silencing Rat - RGC-5 IGF1R

Get tips on using Phusion Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Human - Point mutation A549 ISG15

Get tips on using Rn_Ywhaz_4 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - H9c2 14-3-3 f/Ywhaz

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

Get tips on using siGENOME Rat Lrp5 (293649) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - NPC Lrp5

Get tips on using siGENOME Rat Lrp6 (312781) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - NPC Lrp6