Site Directed Mutagenesis (SDM) Human Point mutation SH-SY5Y

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 ON-TARGETplus Human ARL2 (402) siRNA - Individual to perform siRNA / miRNA gene silencing Human - HeLa ARL2

Get tips on using ON-TARGETplus Human MID1 (4281) siRNA - Individual to perform siRNA / miRNA gene silencing Human - LNCap MID1

Get tips on using ON-TARGETplus Human AR (367) siRNA - Individual to perform siRNA / miRNA gene silencing Human - LNCap AR

Get tips on using ON-TARGETplus Human GAPDH (2597) siRNA - Individual to perform siRNA / miRNA gene silencing Human - H1299 GAPDH

Get tips on using ON-TARGETplus Human AR (367) siRNA - Individual to perform siRNA / miRNA gene silencing Human - DuCaP AR

Get tips on using ON-TARGETplus Human MID1 (4281) siRNA - Individual to perform siRNA / miRNA gene silencing Human - DuCaP MID1

Get tips on using ON-TARGETplus Human GAPDH (2597) siRNA - Individual to perform siRNA / miRNA gene silencing Human - A549 GAPDH

Get tips on using ON-TARGETplus Human PCSK6 (5046) siRNA - Individual to perform siRNA / miRNA gene silencing Human - A253 PACE4