Method for measuring potency of chicken bursin by using in vitro quantitative hemolysis spectrophotometry
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[0026] Preparation of overstocked sheep red blood cell solution (SRBC): Take 5.0ml of sheep venous blood, add it to 5.0ml of Arbor's solution, wash the cells with an appropriate amount of Hank's solution, centrifuge at 1000-2000r / min for 10-20min, discard the supernatant, and wash three times ( The operation is the same as before). Add an appropriate amount of Hank’s solution to resuspend the pelleted cells, centrifuge at 1500-2500r / min for 10-20min, discard the supernatant, and the pelleted cells are the accumulated SRBC cells.
[0027] Preparation of 5% SRBC suspension: take 20 μl of accumulated SRBC, add physiological saline to 400 μl, that is, 5% SRBC suspension.
[0028] Preparation of 10% SRBC suspension: take 20 μl of accumulated SRBC, add physiological saline to 200 μl, that is, 10% SRBC suspension.
[0029] Preparation of 0.18-0.22% sensitized SRBC suspension: take 1ml of normal saline to dissolve the freeze-dried complement, and place it at 4°C for later use; mix th...
Embodiment 1
[0044] Utilize in vitro quantitative hemolysis spectrophotometry to measure the method for chicken bursalin effectiveness, comprising the following steps:
[0045] 1) Preparation of porcine spleen lymphocyte suspension:
[0046] A. Take fresh pig spleen, sterilize it in 75% alcohol, wash it several times with PBS, remove the superficial tissue, cut it into pieces, add Hank's solution to grind it, filter it through a 100-mesh sieve, wash the cells with Hank's solution, 1500r / min Centrifuge for 4 minutes, discard the supernatant, collect the precipitated cells and continue to wash 3 times;
[0047] B. Resuspend the precipitated cells collected after the last wash with an appropriate amount of Hank's solution to obtain a cell suspension, add the cell suspension to a centrifuge tube containing a cell separation medium, centrifuge at 2000r / min for 20min, absorb lymphocytes and place in In another centrifuge tube, add Hank's solution to the centrifuge tube to wash the lymphocytes, ...
Embodiment 2
[0058] Utilize in vitro quantitative hemolysis spectrophotometry to measure the method for chicken bursalin effectiveness, comprising the following steps:
[0059] 1) Preparation of porcine spleen lymphocyte suspension:
[0060] A. Take fresh pig spleen, sterilize it in 75% alcohol, wash it several times with PBS, remove the superficial tissue, cut it into pieces, add Hank's solution to grind it, filter it through a 100-mesh sieve, wash the cells with Hank's solution, 1000r / min Centrifuge for 5 minutes, discard the supernatant, collect the precipitated cells and continue to wash 3 times;
[0061] B. Resuspend the precipitated cells collected after the last wash with an appropriate amount of Hank's solution to obtain a cell suspension, add the cell suspension to a centrifuge tube containing cell separation medium, centrifuge at 1500r / min for 30min, absorb lymphocytes and place in In another centrifuge tube, add Hank's solution to the centrifuge tube to wash the lymphocytes, th...
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