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Get tips on using pIVEX-GAA-omega-PHACTR1-H to perform Protein Expression Eukaryotic cells - N. benthamiana PHACTR1

Get tips on using Napsin A to perform Immunohistochemistry Human - Naspsin A

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Get tips on using M-PER™ Mammalian Protein Extraction Reagent to perform Protein isolation Mammalian cells - SK-N-BE(2)-C

Get tips on using Lectin from Triticum vulgaris (wheat) to perform Western blot 1,4 β-N-acetyl-D-glucosamine - Triticum vulgaris Bacteria Biotin

Get tips on using Anti-NAPSIN A antibody (ab187300) to perform Immunohistochemistry Human - Naspsin A

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.

DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.

Is a knockdown using shRNA permanent and if not is there a known duration?

Get tips on using pET32(a)+-BLS to perform Protein Expression Prokaryotic cells - E. coli BLS