dna-quantification-human-hela

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.

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 Dynabeads™ Untouched™ Human T Cells Kit to perform Cell Isolation Human T cells

Get tips on using MACSprep™ HLA B Cell Isolation Kit, human to perform Cell Isolation HLA B Cell

Get tips on using MACSprep™ HLA T Cell Isolation Kit, human to perform Cell Isolation HLA T Cell

Get tips on using siGENOME Human ATG7 (10533) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - LN-18 ATG-7

Get tips on using siGENOME Human MAPKAPK2 (9261) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - LN-18 MK2/MAPKAPK2

Get tips on using siGENOME Human RIPK3 (11035) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - LN-18 RIP3/RIPK3

Get tips on using siGENOME Human CHUK (1147) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - PANC-1 IKKα/CHUK

Get tips on using siGENOME Human IKBKB (3551) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - PANC-1 IKKβ/IKBKB