Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat schwann cells
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.
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.
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary rat pulmonary artery smooth muscle cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical vein smooth muscle cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical artery smooth muscle cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human renal proximal tubular epithelial cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human pulmonary artery smooth muscle cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human coronary artery smooth muscle cells
Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Cells - primary human hair follicle dermal papilla cells
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