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ELISA experiment setup
ELISA experiment setup
Pre-experimental settings
1. Enzyme linked immunosorbent assay (ELISA) is a biological analysis technique established based on the immunoreaction of antigens and antibodies and the efficient catalytic reaction of enzymes, which can be used to qualitatively and quantify the target protein in the sample to be tested analysis. Common samples to be tested are serum and plasma.
2. For a complete ELISA kits, which contains all reagents, please refer to the instructions directly for the corresponding experimental operations. For the first contact; pre-experiment is a good choice for the ELISA experiment; and for the sample to be tested, the pre-experiment can realize the preliminary judgment of the sample, such as whether the sample needs to be diluted.
3. How to do a pre-experiment?
3.1 First, prepare the sample according to the instructions; if it is not mentioned in the instructions, you need to find the corresponding sample processing method;
3.2 All reagents used should be prepared or prepared according to the instructions. Generally, two strips (16 holes) are set for pre-experiment. Please refer to the instructions for the operation steps. The specific settings of the microplate are as follows:
Take Mouse Brain derived neurotrophic facor, BDNF ELISA Kit as an example, the standard OD values are as follows:
The samples to be tested are brain tissue K1 and M1 (random numbers), choose different dilution multiples for preliminary experiments, and the corresponding ODs are as follows:
Use Curve Expert 1.4 software to draw a standard curve based on the OD and concentration values of the standard, as follows:
Substituting the OD of samples of different dilution multiples into the formula, the corresponding concentration of the sample to be tested can be obtained. (For those with a dilution factor, it needs to be multiplied by the corresponding dilution factor).
The concentration of the sample to be tested is selected to be at a steep curve. For the Mouse Brain derived neurotrophic facor, the BDNF ELISA Kit detects brain tissue, and the stock solution can be considered.
4. Pre-experiment to determine the appropriate dilution factor of the sample, the formal experiment will be carried out below, and the layout is as follows, pay attention to the label, and the formal experiment operation steps please refer to the corresponding manual.
(Note: the competition law is slightly different, you need to strictly follow the instructions)
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