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Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human endothelial cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human mammary mesenchymal cells

Get tips on using NucleoSpin® miRNA to perform RNA isolation / purification Cells - primary human mesenchymal stem cells

Get tips on using Senescence Detection Kit - Biovision to perform Reporter gene assay β-galactosidase substrates - mouse embryonic fibroblasts

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.

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Get tips on using VENTANA anti-MSH2 (G219-1129) Mouse Monoclonal Primary Antibody to perform Immunohistochemistry Human - MSH2

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary canine peripheral blood mononuclear cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human peripheral blood mononuclear cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rat brain microvascular endothelial cells