OptiPrep is the commercial name for an sterile endotoxin solution of 60 percent iodixanol, a chemical originally developed as an X-ray contrast medium. Endotoxins are structural molecules of bacteria, which can be recognized by the body's immune system. Norwegian laboratory Axis-Shield produces this solution, which is widely used in biomedical research, including cellular membrane traffic of substances, as well as virus and bacteria genetics.
Sodium Chloride Protocol
The general purpose OptiPrep sodium chloride gradient preparation using sodium chloride as the osmotic balancer, which keeps the solution's concentration balanced. Technicians make a 50 percent iodixanol working solution by diluting OptiPrep with a standard diluent. They add 0.85 percent sodium chlorine or 0.25 Molar sucrose and Tris-hidrochloride at pH 7.4 to the gradient preparation. According to Axis-Shield , macromolecules have traditionally been purified in gradients containing high concentrations of sucrose, glycerol, alkali metal salts or heavy metal salts. OptiPrep offers the alternative to isolate them under less concentrated solutions.
Protocol for HPV Study
HPV, or human papilloma virus, attacks mucous membranes, causing genital warts and ulcers that can evolve into tumors. The OptiPrep protocol relates to the study of the virus' genetic material, and includes the gradient preparation of OptiPre. Technicians prepare a 46 percent gradient OptiPrep preparation by mixing 70 ml of the OptiPrep, 10 ml of phosphate buffered saline (PBS), 12.5 ml of sodium chlorine, 45 microliters of calcium chloride, 25 microliters of magnesium chloride and 210 microliters of potassium chloride. The preparation must be protected from light and store at room temperature for up to two months.
Isolation of Dendritic Cells Protocol
Dendritic cells are part of the immune system of mammals, often present in tissues close to the skin and in the inner lining of some organs, such as the stomach and intestines. The dendritic cells protocol uses OptiPrep solution in the process of isolating these cells. Technicians mix OptiPrep in a solution of eight percent sodium chloride, Tricine buffer at 7.4 pH and ethylenediaminetetraacetic acid (EDTA). The gradient preparation can be stored at up to 39.2 degrees Fahrenheit for a month.
Tags: gradient preparation, sodium chloride, chloride microliters, immune system, metal salts, OptiPrep solution