Liquid chip kit for diagnosing lung cancer
A liquid phase chip and lung cancer diagnosis technology, applied in the field of medicine and biology, can solve the problems of missed diagnosis, weak specificity, and low sensitivity of patients, and achieve the effect of high accuracy.
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Embodiment 1
[0043] Example 1 Preparation of CTAG antibody, CRP and Prolactin marker detection kit for lung cancer serum samples
[0044] 1. Reagents and solutions
[0045] (1) 0.1M NaH 2 PO 4 , pH6.2: Weigh 3.5814g NaH 2 PO 4 in 90mL ddH 2 In O, adjust the pH to 6.2 with NaOH, dilute to 100mL, and filter with a 0.22um filter;
[0046] (2) 0.05M MES, pH5.0: weigh 0.976g MES in 90mL ddH 2 In O, adjust the pH to 5.0 with NaOH, dilute to 100mL, and filter with a 0.22um filter;
[0047] (3) 0.1M MES, pH6.0: weigh 1.952g MES in 90mL ddH 2 In O, adjust the pH to 6.0 with NaOH, dilute to 100mL, and filter with a 0.22um filter;
[0048] (4) 0.1M MES, pH4.5: weigh 1.952g MES in 90mL ddH 2 In O, adjust the pH to 4.5 with NaOH, dilute to 100mL, and filter with a 0.22um filter;
[0049] (5) PBS: NaCl, 137mM; KCl, 2.7mM; NaCl 2 HPO 4 , 8.1mM; KH 2 PO 4 , 1.5mM; pH7.2-7.4, 0.22um membrane filtration;
[0050] (6) 1%PBSB: 0.1%PBS containing 0.02%Proclin300, 0.22um membrane filtration;
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Embodiment 2
[0100] Embodiment 2 Liquid phase chip kit of the present invention is used for the detection of lung cancer
[0101](1) Take out all the reagents before use, and let them equilibrate to room temperature;
[0102] (2) Take 2-plex Halo Tag (O 2 ) Amine ligand-coupled microspheres, vortexed for 20s, ultrasonicated for 20s, according to 2500 microspheres per well, take an appropriate amount in 500ul of 1%PBSB;
[0103] (3) Protect from light and seal for 1 hour;
[0104] (4) 14000g, centrifuge for 4min, absorb the supernatant and add the corresponding protein, among them, CTAG (Creative Biomart, Cat. No. CTAG1B-1512H), Halo each 0.2ug / well, supplemented with 1% PBSB to a final volume of 500ul;
[0105] (5) Incubate at room temperature in the dark for 1 hour;
[0106] (6) After vortexing and ultrasonicating the above microspheres for about 20s, dispense them into 96-well plates, 10ul / well;
[0107] (7) Prepare CRP (R&D Systems, product number 1229006), Prolactin (R&D Systems, p...
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