A kind of Alzheimer's disease detection reagent and its application in Alzheimer's disease detection
A technology for Alzheimer's disease and detection reagents, which is applied in the field of biomedicine to achieve the effect of high repeatability and good stability
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Embodiment 1
[0039] (1) Experimental equipment:
[0040] ABI3500dx analyzer (Applied Biosystems), PCR, centrifuge.
[0041] (2) Reagent preparation:
[0042] AD1: Mix A and B 1:1 by volume.
[0043] A: SDS (ammonium dodecyl sulfate) was dissolved in 10mM NH4HCO3, and prepared as a solution containing 5% SDS and pH 8.3
[0044] B: Mix 2.2U / µL PNGaseF and 3.33% NP-40 at a volume ratio of 1:20
[0045] AD2: Mix the same volume of 20mM APTS and 1M NaCNBH3.
[0046] AD3: 100mM NH4AC, sialidase (neuraminidase, 2mU / µL) and hydrogen peroxide according to volume 5:1:14
[0047] mix.
[0048] AD4: buffer.
[0049] (3) N-oligosaccharide chain map detection
[0050] a. Preparation of oligosaccharide chains: Take 2 µL of serum inactivated at 95°C, add 4 µL of AD1, and react at 37°C
[0051] Dry after 3h.
[0052] b. Labeling of oligosaccharide chains: Add 2 µL of reagent AD2 to the dried sample at 65°C
[0053]After 3 hours of fluorescent labeling, 200 µL of AD4 was added to terminate the rea...
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