Detailed explanation of ancient creatures: washing method of Elisa experiment

Detailed explanation of ancient creatures: washing method of Elisa experiment

Elisa experimental washing method In addition to some ELISA instruments equipped with a special automatic washing machine, manual operation has two types of immersion and running water, the process is as follows:

(1) Immersion type:

a. blot or dry the reaction solution in the well; b. wash it once with the washing solution (after filling the plate with the washing liquid, it will be removed);

c. Soaking, the washing liquid is filled into the plate hole, placed for 1-2 minutes, intermittent shaking, the soaking time can not be shortened arbitrarily;

d. Drain the liquid in the well. Drying should be thorough, can be pumped by a water pump or a vacuum pump, or can be patted on a clean towel or absorbent paper after removing the liquid;

e. Repeat operations c and d, wash 3-4 times (or as specified). In the indirect method, if the background is higher, the number of washings or the soaking time can be increased.

Microtiter plates are mostly immersed. The washing solution is mostly a neutral buffer containing a nonionic detergent. The binding of the polystyrene carrier to the protein is hydrophobic. The nonionic detergent contains both a hydrophobic group and a hydrophilic group, and the hydrophobic group binds to the hydrophobic group of the protein through a hydrophobic bond, thereby weakening the protein and The solid phase carrier is combined, and by the combination of the hydrophilic group and the water molecule, the protein is returned to the aqueous solution state, thereby being separated from the solid phase carrier. The non-ionic detergent in the washing liquid is generally Tween 20, and its concentration may be between 0.05% and 0.2%. When the concentration is higher than 0.2%, the antigen or antibody coated on the solid phase may be desorbed to reduce the test. Sensitivity.

(2) The running water rinse type water washing method is originally used for the washing of the bead carrier, and the washing liquid is only distilled water or even tap water. A special device is attached during washing to continuously roll and wash the beads under the impact of running water. After rinsing for 2 minutes, the liquid is drained, soaked in distilled water for 2 minutes, and blotted dry. The immersion is like a bath, and the running water is like a shower. The washing effect is more thorough and simple and fast. It has been shown in experiments that the flow rinse type is also suitable for the washing of microtiter plates. Try to increase the water flow or increase the water pressure during washing, so that the water flow impacts the surface of the plate hole, and the washing effect is better.

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