rna-isolation-purification-cells-immortalized-ags

Get tips on using psiCHECK-2vector to perform Reporter gene assay luciferase - HEK 293 human embryonic kidney cells

Get tips on using pGL3-Basic Vector to perform Reporter gene assay luciferase - vascular smooth muscle cells (VSMC)

Get tips on using CometChip Electrophoresis Starter Kit to perform DNA Damage Assay Human bronchial epithelial cells (hBE)

Get tips on using CellROX™ Deep Red Reagent, for oxidative stress detection to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

Get tips on using Beta-Glo® Assay System to perform Reporter gene assay β-galactosidase substrates - rat mesenchymal stem cells (MSCs)

A restriction enzyme or restriction endonuclease is defined as a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at or near that site, known as restriction site or target sequence. The four most common types of restriction enzymes inclue: Type I (cleaves at sites remote from a recognition site), Type II (cleaves within or at short specific distances from a recognition site), Type III (cleave at sites a short distance from a recognition site), and Type IV (targets modified DNA- methylated, hydroxymethylated and glucosyl-hydroxymethylated DNA). The most common challenges with restriction digest include- 1. inactivation of enzyme, 2. incomplete or no digestion, and 3. unexpected cleavage. The enzyme should always be stored at -20C and multiple freeze-thaw cycles should be avoided in order to maintain optimal activity. Always use a control DNA digestion with the enzyme to ensure adequate activity (to avoid interference due to high glycerol in the enzyme). For complete digestion, make sure that the enzyme volume is 1/10th of the total reaction volume, optimal temperature is constantly maintained throughout the reaction, the total reaction time is appropriately calculated based on the amount of DNA to be digested, appropriate buffers should be used to ensure maximal enzymatic activity, and in case of a double digest, make sure that the two restriction sites are far enough so that the activity of one enzyme cannot interfere with activity of the other. Star activity (or off-target cleavage) and incomplete cleavage are potential challenges which may occur due to suboptimal enzymatic conditions or inappropriate enzyme storage. To avoid these, follow the recommended guidelines for storage and reactions, and always check for the efficacy of digestion along with purification of digested products on an agarose gel.

Get tips on using Pierce™ BCA Protein Assay Kit to perform Protein quantification Mammalian cells - MDA-MB-231

Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Mammalian cells - MDA-MB-231

Get tips on using Pierce™ Coomassie (Bradford) Protein Assay Kit to perform Protein quantification Mammalian cells - Mouse macrophages

Get tips on using Pierce™ Coomassie (Bradford) Protein Assay Kit to perform Protein quantification Mammalian cells - Neuro 2a