ELISA kit production process and precautions

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1. ELISA kit production process

2, the coating

2.1 Prepare the coating solution with the coating antigen in the coating buffer , add 100 m L per well to the enzyme labeling plate, cover the cover film, and coat the conditions as shown in the following table. The coating plate can be coated at 4 °C for 16-20h. Stacked, and the spacing between the plates should be consistent (typically 5 cm ) when coated at 37 °C for 2 h . According to the design of the incubator, the spacing between the microplates is adjusted. If the incubator generates heat from the bottom, the spacing between the lower microplates should be increased to 10 cm .

2.2 packets are loaded to take two dozen withdrawing a manner to avoid the hole wall portion is applied, if inadvertently dropped on the pore walls, a final decision should be based on whether the solution was dropwise added to the wells, if added should, carefully Shake it to the bottom of the hole, otherwise use a suction paper to carefully suck out , and be careful not to spill, no bubbles.

2.3 Pre-sucking before the bag is checked to see if the gun head is qualified, the water flow is consistent, the inconsistency should be replaced, the bag should be made of high quality gun head, each bag should be a plate, the gun head should be tightly clamped , and in the process of coating Pay attention to check whether the aspiration is consistent. If there are any minor problems during the coating process, the board should be marked so that it can be eliminated when the group is loaded into the library.

2.4 If the coating mode is used at 37 °C for 2 h , the enzyme labeling plate should be coded to ensure that the coating time of each plate is consistent.

3 , wash the board

3.1 After the coating, two washings should be carried out, 250 μl / well. After washing, the liquid should be patted on the towel and the next step should be closed in time.

3.2 When washing, pay attention to check whether the water flow is consistent. If any small problems occur during the washing process, the board should be marked so that it can be eliminated when the group is loaded into the library.

4 , closed

4.1 Blocking solution shall be taken one hour before initial warm, shake well before use, take each well was added 1 8 0 m L ELISA plate, cover plate film, the spacing between the closure 3 7 ℃ 1.5h, microtiter plates Should be consistent (typically 5cm ). According to the design of the incubator, the spacing between the microplates is adjusted. If the incubator generates heat from the bottom, the spacing between the lower microplates should be increased to 10 cm .

4.2 Closed sample loading should be avoided by adding a dozen. If it is accidentally dropped on the wall of the hole, it should be judged according to whether the solution should be added to the hole. If it should be added, carefully shake it. Let it fall into the bottom of the hole, otherwise use a suction paper to carefully suck out , and the solution remaining in the tip of the gun should not be shot to avoid air bubbles. And pay attention to not spilling, no bubbles

4.3 Pre-sucking before closing to check whether the gun head is qualified, whether the water flow is consistent, the inconsistency should be replaced by the gun head, and the high-quality gun head should be used for closing. The gun head should be tightly closed for each closed board , and pay attention to the inspection during the coating process. If the aspiration is consistent, if there are any minor problems during the sealing process, the board should be marked so that it can be eliminated when the group is loaded into the library.

5 , drying bagging

5.1 After closing, drop the liquid in the hole and pat dry on the towel, then dry or dry the enzyme plate thoroughly. Drying: 37 ℃ inversion (or uncovered by the plate film ziplock), each plate 5 Drying 30 min, with a corresponding increase in the number of the extension board time, such as board 100, needs to bake 3 h, and so on sequentially; Dry: Put a layer of clean A4 paper or absorbent paper on the bottom , invert the board, cover it with a layer on the top to protect it from light, and let it cool for 18-24 hours at room temperature .

5.2 After the board is dried, it should be loaded into the ziplock bag in time, and filled in according to the amount of one bag of desiccant per plate. The date of preparation of the board is written on the outside of the bag, and stored in the refrigerated environment of the finished product warehouse for storage.