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 Lipofectamine® 2000 Transfection Reagent to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells hsa-miR-542-3p Lipid
Get tips on using Silencer® Select GLO-1 siRNA to perform siRNA / miRNA gene silencing Human - Primary Human Aortic Endothelial Cells GLO-1 Lipid
Get tips on using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Human - Deletion K562 c-Myb gene
Get tips on using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - HepG2 FHIT
Get tips on using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - Hep3B SFRP3
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 EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - TCP-1, BCPAP & nthy-ori 3-1 (thyroid tumor cells) METTL7A
Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - TCP-1, BCPAP & nthy-ori 3-1 (thyroid tumor cells) BCPAP
Get tips on using β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer to perform Reporter gene assay β-galactosidase substrates - human MSCs (mesenchymal stem cells)
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