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Although washing is not a reaction step during the ELISA test, it also determines the success or failure
of the experiment.
The ELSIA test is done by washing to separate free and bound enzyme markers
.
By washing to remove the remaining substances in the plate wells that fail to bind to solid-phase antigens or antibodies, as well as interfering substances
that are non-specifically adsorbed to solid-phase carriers during the reaction.
The adsorption of proteins by plastics such as polystyrene is universal, and this non-specific adsorption interference substance should be washed down
when washing.
It can be said that in the ELISA test operation, washing is the most important key technology
.
In addition to some ELISA test instruments equipped with special automatic washing instruments, manual operation has two types: immersion type and running water flushing type.
The process is as follows: 1.
The soaking type (1) blotting or drying the reaction liquid in the hole; (2) washing it once with the washing liquid; (3) soaking, that is, filling the plate well with the washing liquid, placing it for 1 to 2 minutes, shaking intermittently, the soaking time cannot be shortened at will; (4) the liquid in the suction hole, the suction should be thorough, can be suction with a water pump or vacuum pump, and can also be thrown off the liquid and patted dry on the cleaning towel; (5) repeat the operation (3) and (4), wash 3 to 4 times
.
In the indirect method, if the background is higher, it can increase the number of washes or extend the soaking time
.
2.
The flowing water flushing method was originally used for the washing of the bead carrier, and the washing liquid was only distilled water or even tap water
.
A special device is attached to the washing, so that the beads are continuously rolled and rinsed under the impact of running water, and after continuous rinsing for 2 minutes, the liquid is sucked dry, and then soaked in distilled water for 2 minutes, and it can be
sucked dry.
The soaking type is like a bathtub, and the running water flushing type is like a shower, and its washing effect is more thorough, and it is also simple and fast
.
Experiments have shown that the flowing water flushing formula is also suitable for the washing
of microtiter plates.
When washing, try to increase the water flow or increase the water pressure, so that the water flow hits the surface of the plate hole, and the washing effect is better
.