2 English reference APOB
Apolipoprotein b
Apolipoprotein is a part of plasma lipoprotein, and all kinds of lipoproteins contain one or several different specific apolipoproteins. At present, more than 20 kinds of apolipoproteins have been found. ApoB is one of them.
APOB, another name for apolipoprotein B.
5 Apolipoprotein B medical examination 5. 1 examination name Apolipoprotein B.
5.2 Classified blood biochemical examination/blood lipid determination
5.3 Test blood samples
5.4 Principle The principle of apolipoprotein B is that when antigen and antibody react in a certain proportion, fine particles of antigen-antibody complex are generated in the solution and uniformly dispersed in the solution medium. When light passes through this turbid liquid, the particles in the turbid liquid can absorb light, and the amount of absorbed light is proportional to the amount of turbid particles. Under certain conditions, serum ApoA 1 or ApoB combines with corresponding antibodies to form insoluble immune complex, which makes the solution turbid, that is, turbidity is directly proportional to absorbance.
5.5 reagent (1) barbital buffer, ionic strength 0.05, pH8.6.
(2) prepare 10g/L agarose (standard electroosmosis) with barbital buffer, heat and melt, mix, sub-package to 10ml/ tube, cool and store at 4℃.
(3) 0. 1.5 mol/l sodium chloride solution.
(4) rabbit (or sheep) anti-human apoA antiserum (titer not lower than 1: 16) and apoB antiserum (titer not lower than 1: 32).
(5) Calibration serum (year-on-year turbidity method).
(6) Dyeing solution: dissolve 0.25g of Coomassie Brilliant Blue R250 in 45ml of methanol, add glacial acetic acid10ml and 45ml of water, and mix and dissolve for later use.
(7) Decolorization solution: each liter contains 50 ml glacial acetic acid and 100 ml glycerol.
5.6 operation method (1) pour plates: each plate is melted in boiling water bath with 10g/L agarose 10ml, and mixed evenly. When it is cooled to 50 ~ 55℃, add 50μ l of apoa I antiserum and 8μ l of apob antiserum (the dosage depends on the titer of antiserum) and mix quickly. Put the plate into the electrophoresis tank for use.
Filter paper bridging
(2) antigen dilution: dilute the calibration serum to 1: 100, 1: 150, 1: 200, 1: 300 with 0: 05 mol/L NaCl solution.
(3) Sample addition: Under the low current state (10mA/ plate), 5μl of diluted serum and sample were respectively absorbed (accurately) and added into the agarose gel sample addition hole. Each committee must set a series of standards.
(4) electrifying: steady flow of 24mA/ plate, terminal voltage of 6 ~ 8 V/cm, running water cooling to keep agarose at 65438 05℃, and electrophoresis for 3 ~ 4 hours.
(5) Removing impurities and drying the adhesive film: soak the electrophoresis agarose plate in 0. 1.5 mol/L NaCl for 30 minutes, put the adhesive film on the polyester film, absorb the moisture in the adhesive with multilayer filter paper under slight pressure, then naturally dry the filter paper, adhesive film and polyester film together, or blow dry with hot air, and the dried adhesive film will naturally separate from the filter paper and polyester film.
(6) Dyeing: the agarose film is laid flat and soaked in the dyeing solution for 20-30 minutes.
(7) Decolorization: soak the dyed film in decolorizing solution until the rocket peak is clear and the background is basically colorless. The film can be sandwiched in water with two pieces of cellophane, and can be stored for a long time after drying. You can also use running water to soak and decolorize until the background is clean.
5.7 Normal value: apoa1:1.00 ~1.50g/l; Apolipoprotein b:0.50 ~ 1. 10 g/l.
5.8 Clinical Significance of Laboratory Results ApoA 1 level is negatively correlated with hyperlipidemia and coronary heart disease. ApoB level is positively correlated with hyperlipidemia and arteriosclerosis.
5.9 Note (1) ApoA 1 and ApoB antiserum are purchased with high titer and high specificity.
(2) In order to accurately determine ApoA 1 and ApoB, a standard curve must be made.
(3) The immune transmittance should be calibrated at multiple points. Perform regression operation according to the curve.
5. 10 hyperlipidemia-related diseases