Do you do the right thing in monoclonal antibody preparation, blood cell purification, and organelle separation?
General purpose centrifuges are used in a wide range of industries. At present, the centrifuge products used in scientific research units are high-speed centrifuges, blood centrifuges, cell centrifuges, etc., with a wide variety of functions and functions. Different experiments require different centrifugation conditions, and it is often necessary to centrifuge the cell suspension to increase the cell concentration or to wash away the relevant reagents. The adjustable tabletop centrifuge can meet the needs of most experiments, so this article briefly introduces the preparation of the monoclonal antibody. How to use the centrifuge properly in the three experiments of blood cell purification and organelle separation.
Monoclonal antibody preparation
Monoclonal antibody technology An immunological technique in which a single B lymphocyte producing an antibody is hybridized with a bone marrow tumor cell to obtain a hybrid cell which can produce both an antibody and an infinite proliferation, and thereby produce an antibody. This technology is widely used in disease diagnosis, treatment and test diagnosis in many industries. Each step of antibody preparation requires the use of a centrifuge. Reasonable centrifugation conditions are an effective guarantee for obtaining monoclonal monomers.
1. There may be a variety of immunogens: cells, cell extracts, purified proteins, fusion proteins, polypeptides, carbohydrates, lipids or DNA. When the cells are used as immunogens, the rotation speed should be 600 rpm/min~1000 rpm/min, and the temperature should be centrifuged for 5 min~10 min at room temperature. If the rotation speed is too low, the cell sedimentation will be too small, and the rotation speed will cause cell breakage. The purified protein and the fusion protein should be used as an immunogen at a speed of 10,000 rpm/min to 15000 rpm/min and centrifuged at 4 ° C for 20 to 45 minutes.
2. In the process of cell fusion, cell screening and cell cryopreservation, the speed is 600 rpm/min~1000 rpm/min, and the temperature can be centrifuged for 5 min~10 min at room temperature.
Blood cell purification
Blood cells are composed of many types of cell suspensions, each with its own characteristics and functions and widely used in medical clinical and diagnostic applications. Researchers have done a lot of work on blood cell separation and purification for many years, and various centrifugal equipment have been widely used in laboratory research and blood component separation. Below we will use laboratory research as the main line to illustrate the various blood cell separation and centrifugation conditions:
1. Mononuclear (white) cells need to be purified from 1800 rpm/min to 3000 rpm/min and centrifuged at 20 °C for 15 min to 20 min (do not centrifuge at 0~5 °C, otherwise the separation will be poor). It can be used at 1000 rpm/min ~ 1300 rpm/min during the washing process and centrifuged at 20 ° C for 5 min.
2, human blood separation of red blood cells, mononuclear (white) cells, neutral (white) cells need 1500 rpm / min ~ 1600 rpm / min, 18 ° C ~ 23 ° C centrifugation 35 min, the temperature is too high during centrifugation, centrifugal force Large or centrifugation for too long may cause neutral (white) cells to precipitate.
Organelle separation
Organelle separation is the basic means of studying such as nucleus, mitochondria, Golgi, lysosomes and microsomes, as well as various macromolecules. The tissue is usually homogenized using various physical methods such as grinding, sonication, hypotonicity, and then the organelles are obtained by differential centrifugation and density gradient centrifugation. Let us briefly understand the centrifugation conditions of different organelles:
The Esco Versati range of centrifuges is equipped with maintenance-free motors, robust mechanisms and a smart Ecourage microcomputer processor control system for maximum reliability and safety. Covering microcentrifuges and low-speed universal centrifuges, it has many types of rotors, adapters and accessories. It is suitable for a variety of general-purpose tubes, tubes and microplates, with a maximum speed of 16000 rpm. Range -20 ° C ~ 40 ° C.
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