Separation of higher apolipoprotein-containing components (HDL, VHDL) by electrophoresis was successful, while centrifugation was suitable for separation of serum lipoproteins of various densities. Serum lipoproteins are a heterogeneous complex of lipids in the blood that are composed of certain proteins. Due to the difference in the ratio of lipid to apolipoprotein, the density ranges from 0.96 g/ml or less to 1.21 g/ml. Paper electrophoresis shows four bands: milk dense particles, very low density lipoprotein, low density lipoprotein, high density lipoprotein. Each lipoprotein can also be divided into more sub-components. one. Gradient material: The serum lipoprotein density is low. The commonly used gradient materials are Nacl, NaI, Kbr, KI, etc. The commonly used buffer is ED. two. Pre-separation: 1. Separation of blood cells from serum: Centrifugal parameters: whole blood, angular head, 3,000 rpm × 20 min Results: The precipitate was blood cells and the upper part was serum. 2. Separation of chylomicrons: The chylomicron content in human plasma is very low after fasting for more than 12 hours. Those who have eaten have high chylomicron in their plasma, and should first centrifuge to remove chylomicron. Centrifugal parameters: 4 × 10 (g × min), 10 ° C. Various rotor speeds can be. Gradient solution configuration: 3/4 volume of the lower part of the centrifuge tube plus plasma, upper 1/4 volume plus 0. 5MnaCl 0.3MEDTA, ph7.4 Result: The chylomicrons floated up and sucked out. 3. Separation of serum proteins: removal of globulin, albumin and other proteins. Centrifugal parameters: (2.5 to 3) × 10 (g × hr), all kinds of rotors can be. Such as angle rotor 70,000 rpm × 6hrs, 10 ° C Configuration: tube capacity 1/3 is serum, 2/3 is 1.31g/ml, NaCl NaBr, the final density after stirring is 1.21g/ml Results: 1/6 of the upper volume of the tube was serum lipoprotein, and the lower 5/6 was other proteins. three. method: 1. Sequential flotation method: a) Overspeed method: the earliest method used, 4000 rpm × 24 hrs × 3 times, 10 ° CNaCl NaBr gradient according to 1.006, 1.063, 1. 21 sequential flotation, each time taking different density in the upper part of the tube lipoprotein. The operation is simple, the resolution is high, the yield is high, time-consuming and costly. b) Ultra-high speed method: 100,000 rpm × 2.5 hrs × 2 times, 10 ° C, 100,000 rpm × 4 hrs × 1 time, NaCl KBr EDTA gradient flotation VLDL, LDL, HDL without density 2. Single centrifugation: a) Single horizontal separation The maximum speed is 25,000~28,000 rpm, and the 6×40ml flattening head is configured with 1.006g/ml NaCl solution and 1.35g/ml (NaCl KBr EDTA) solution to form 8 layers from 1.006 to 1.21. Do not Continuous step, the lowermost serum was combined with KBr to form 1.25g/ml sample solution, 25,000 rpm×4hrs, and VLDL, LDL, HDL different layer zones were obtained once at 4oC. With a maximum speed of 40,000 rpm, 6 × 13ml 甩 flat head, 40,000 rpm × 24hrs, 20 ° C, discontinuous KBr gradient 1.006 ~ 1.21g / ml, slender tube, clear layer, recovery High, long centrifugation time b) Single vertical tube separation: The NaCl/KBr or NaCl/NaBr discontinuous gradient single tube capacity is from 5ml to 40ml, the rotation speed is from 50,000rpm to 80,000rpm, centrifugation (0.5-3)hrs, and the lower layer is adjusted to 1 by KBr or (NaBr). . The upper layer of 3g/ml was added with 1.006g/ml, NaCl solution, 10°C-20°C, and decelerated once. The NaCl/Sucrose discontinuous gradient was 48% W/WSucrose in the lower layer and the middle serum was in 4MnaCl. In the middle layer, the upper layer is 0. 67MNaCl 0.05% EDTA, and the centrifugation parameters are the same as above, and the time is longer. c) Zone turret large capacity separation: Centrifugal parameters: zone rotor, maximum speed 30,000~48,000rpm, capacity from 650ml~1900ml, serum 50-150ml can be separated each time, low speed loading and unloading, separation speed: big turn (25,000-28,000 rpm) × 24 hrs, small turning head (30,000-38,000 rpm) × 1 hrs, 10 ° C - 20 ° C Gradient: NaCl NaBr or KBr EDTA is discontinuous or linear gradient. The sample is in the maximum density layer, and the pure core is used as the isolation layer at the core of the near-head. The outermost part of the separation chamber is made of high-density NaCl/KBr or NaCl/NaBr was used as a buffer layer. General Surgery Table,General Operating Table,Hydraulic Operating Table,Mechanical Hydraulic Operating Table NINGBO TECHART MEDICAL EQUIPMENT CO.,LTD , https://www.techartmed.com