Method for determining minim proteins based on magnetic pearl and nano gold probe
A nano-gold probe and assay method technology, which is applied in the field of protein assay, can solve the problems of failing to meet the clinical detection requirements, lower detection sensitivity, and complicated operating procedures, and achieve the effects of high sensitivity, convenient operation, and fewer operating steps
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[0035] Using this technology to analyze human p53 positive serum, the p53 protein content is 160pg / ml.
[0036] 1P53 Monoclonal Antibody Labeled Magnetic Beads
[0037] 1.1 Activate magnetic beads: Wash 300 μl magnetic beads (equivalent to 3 mg magnetic beads) twice with 300 μl 25mM MES (pH6), add 50 μl of 50 mg / ml EDC solution and 50 μl of 50 mg / ml NHS solution in ice bath, and incubate at room temperature for 30 minutes; Wash twice with 300 μl 25 mM MES (pH 6).
[0038]1.2 Labeling: Add 60 μl of p53 antibody, add 40 μl of 25mM MES (pH6), to make the final volume 100 μl; place at 4°C for 2 hours; wash 4 times with 300 μl of PBS (containing 0.1% BSA and 0.01% Tween-20), and then Resuspend the magnetic beads with 300 μl washing solution and store at 2-8°C for future use.
[0039] 2p53 Polyclonal Antibody and DNA Probe Labeling Nanogold
[0040] 2.1 Determine the optimal amount of antibody. Use 0.1M K2CO3 solution to adjust the pH value of the nano-gold solution to 9, and tak...
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