Method for testing BNP (Brain Natriuretic Peptide) activity by using GCA stable transfection cell line
A technology of transgenic cells and reactivity, which is applied in the measurement/inspection of microorganisms, cells modified by introducing foreign genetic materials, biochemical equipment and methods, etc., can solve the problems of large errors and cumbersome operations, and achieve small variation and high efficiency. Application value, high accuracy effect
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[0044] 1. Materials and methods:
[0045] 1.1 Research object: B-type brain natriuretic peptide (BNP)
[0046] 1.2 NPR1 / GCA plasmid: the gene ID is NM 000906, purchased from Aurui Dongyuan Gene Co., Ltd., see the vector map image 3 .
[0047] HEK293 cells: stored in the cell room of China Food and Drug Research Institute.
[0048] BNP reference product: prepared by the Recombinant Technology Product Office of China Institute of Food and Drug Research and Testing.
[0049] 1.3 Reagents:
[0050] Growth medium: DMEM (GIBCO, #11995) + 10% fetal bovine serum (GIBCO, #10099) + 1% double antibody (GIBCO, #15240)
[0051] Plate medium: DMEM (GIBCO, #11995)
[0052] Liposome 2000: Invitrogen, #11668
[0053] G418: CalBiochem, #345810
[0054] G418 selection medium: growth medium + 200ug / ml G418
[0055] BNP diluent: PBS+0.2%BSA (Merck Calbiochem, #12659)+1mM IBMX (sigma, I5879, add just before use)
[0056] cGMP detection kit: cell signaling, #4360S
[0057] cGMP primary an...
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