Analysis of common problems in Elisa experiment - Database & Sql Blog Articles

EL-C1600N100013-B

SMD aluminum electrolytic capacitor

Brand AVX TPSE226M035R0125 Low impedance tantalum capacitor AVX 22

Probe domestic double-head probe 038-BB-5.7L The shape of the two ends of the needle is outside the pointed needle

When we do experiments, we often encounter

The problem of low color sensitivity of the kit is low. The following is the year-end summary of the senior technology of Shanghai Jinma Biotechnology Co., Ltd. based on the previous years. According to the different reasons encountered in the personal experiment and the possible reasons and their prevention methods, for reference only:

1

. Insufficient incubation time

Fixed clock timing

2

. The impact force during washing is too large, the soaking time of the washing liquid is too long, and the washing time is increased.

Reduce washing impact, keep washing time according to the instructions, and accurately remember the washing times

3

. Insufficient amount of developer added or reversed, or added after mixing

When the coloring solution is added dropwise, the dropping bottle is vertically downward, the holding force is uniform, and the dropping speed should not be too fast, first adding the coloring agent.

, anti-additive coloring agent

Can't

,

Add after mixing

4

. Insufficient reaction time of the developer

Accurate timing

5

. There is a problem with the quality of distilled water

Determination of the effect of distilled water preparation reagents on enzyme immunoassay

6

. The kit is too long in transit and the temperature is too high

The kit is transported in an incubator and a sufficient amount of ice is placed to minimize transit time.

7

. Reagents and samples failed to balance before use

Pre-reagents and samples are equilibrated at room temperature

Minutes or so.

8

. Sample and

Antiseptic, inhibits the reaction of the enzyme

Sample cannot be added

Antiseptic

9

. The kit is over the expiration date

Do not use more than the expiration kit

10

. Reagent opening time is too long

Please try to use up in a short time after the reagent is turned on.

11

. The pipette is not enough, the pipetting suction is too fast, the liquid on the inner wall of the tip is too much or the inner wall is not clean.

Correct the pipette, the tip should be matched, and the tip should be tightly matched each time. The pipetting should not be too fast and the discharge should be complete. The inner wall of the tip should be clean and preferably used at one time.

12

. Insufficient temperature in the oven

°C or water bath pot water temperature is insufficient

°C)

When placing the reaction plate, pay attention to the temperature of the incubator and adjust it in time. It should not be too much during the incubation period to affect the temperature balance.
The ELISA assay modes include the following: double antibody sandwich method, indirect method, double antigen sandwich method, IgM antibody capture method, competitive inhibition method, in which the competition inhibition method (HBeAb, HBcAb, etc.) is unfair due to the operation time difference. The impact of factors such as competition, the results are less repeatable, and the quality is more difficult to control.
Different batches of ELISA reagents are difficult to ensure the same quality during the production process. Even if the results are different through batch inspection, it is necessary to select and order long batch reagents and ensure storage conditions. Strict implementation of this standard can avoid the complicated process of re-establishing the quality control system and re-evaluating the reagents due to the change of the reagent batch number, and can ensure the stability of the results; for reagents with short pot life and low usage rate, it should be dispensed in small quantities. For each use, take the dispensing part to avoid the failure of the reagent caused by repeated freezing and thawing.

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