ELISA reagent kit for on-site detection
An on-site detection and kit technology, applied in biological testing, material inspection products, etc., can solve problems such as inconvenience in carrying and mailing, failure of kits, impact of antibody and enzyme titers, etc., to increase biological safety and increase resistance. The effect of increasing the range and seismic performance
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Embodiment 1
[0083] Preparation and use of TAS-ELISA detection kit for Cymbidium mosaic virus (CyMV) on-site detection. The kit contains ①antibody detection plate, ②sample dilution reagent, ③washing reagent, ④quality control (positive / negative control), ⑤ enzyme conjugate reagents (including detection antibodies, enzyme-labeled anti-antibodies, diluents), ⑥ substrate reagents, ⑦ termination reagents, ⑧ some auxiliary equipment.
[0084] 1. The specific preparation process is as follows:
[0085] 1.1 Antibody detection plate:
[0086] Preparation of coating buffer, containing Na 2 CO 3 1.59g / L, Na 2 HCO 3 2.93g / L, NaN 3 0.2g / L to adjust the pH to 9.6. Take 50 microliters of coating antibody (rabbit anti-Jianlan mosaic virus, produced by American company Agdia) and dilute it 1:200 times with coating buffer (that is, add 9.95 mL of coating buffer per 50 μL to prepare 10 mL of use solution) , on a detachable 96-well high-affinity microtiter plate (produced in Canada), add 100 microlite...
Embodiment 2
[0174] TAS-ELISA detects the preparation and use of CyMV on-site detection kit, the difference with embodiment 1 is only described below: the carrier used is a cotton ball; the detection antibody does not contain an indicator, and an indicator is added on the enzyme-labeled anti-antibody Reagent I; The indicator I used is acid chrome blue K; The indicator II is preserved in a small centrifuge tube.
[0175] 1.5.2 Preparation of temporary solid carrier for antibody: Dissolve 0.05 g of Guangming brand skimmed milk powder in 10 ml of enzyme conjugate dilution buffer → knead the absorbent cotton into small balls of 3-4 mm (mung bean size), soak for 2 hours, during each Stir once every 30 minutes → soak in washing buffer 4-6 times → control the water, spread it on a clean glass slide, dry it in a desiccator, and store it in a desiccator for later use. Thereby obtaining a temporary solid carrier of the antibody.
[0176] 1.5.3 Preparation of absorbent cotton pellets containing dete...
Embodiment 3
[0183] DAS-ELISA detection of Potato Leaf Roll Virus (Potato Leaf Roll Virus) (antibody produced by agdia company) field detection kit preparation and use. Only the difference from Example 1 is described below: 1. The enzyme conjugate reagent only contains enzyme-labeled antibody and enzyme conjugate diluent; 2. The indicator is added to the enzyme conjugate diluent, and the indicator used is methyl red.
[0184] 1. Prepare enzyme conjugate diluent: weigh Na 2 HPO 4 0.620g, KH 2 PO 4 0.100g, and NaCl 4.00g, KCl 0.10g, NaN 3 0.10g, bovine serum albumin 1.0g, polyvinylpyrrolidone K3010±1g, Tween -20 0.25±0.05g→dissolve reagent in 80ml pure water→add 1mL of 1g / L methyl red indicator, mix well→use 0.10g / mL Na 2 HPO 4 solution; or 0.02g / mL KH 2 PO 4 Adjust the pH of the solution to 7.03→concentrate to 100mL to obtain a 5-fold concentrated solution of ECI→test: take 2mL and dilute it with water to 10mL to obtain the enzyme conjugate dilution buffer (pH 7.40~7.42)→fill e...
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