Introduction:
ELISA stands for enzyme-linked immunosorbent assay, also known as an enzyme assay, is used to test for many diseases by detecting either the antibody or the antigen. ELISA is a powerful diagnostic tool in human medicine, disease detection, animal, plants, detecting illegal drug used, and testing indoor air quality and food safety. A sample contains many proteins and may or may not contain the antigen, the two unknown antibody will bind to the antigen if presence in a sample, the last was to evaluate the assay results after enzyme substrate added to the wells. Positive and negative controls are used to compare to the test well. The positive control has a strong color change; the negative control has little or no color change.
Materials: Micro plate, multi channel pipette, single channel pipette, micropipette tips, tips disposable cup, ELISA wash buffer, paper towels, reagent reservoir, positive control, negative control, primary antibody (1°Ab), secondary antibody (2°AB), enzyme substrate, unknown sample A and sample B.
Methods: Labeled micro plate first row 12 wells in order with ++--AAAABBBB. Used new tip with each type of sample, loaded 50μl from positive sample to + wells, negative sample to – wells, A sample to A wells and B sample to B wells labeled. Waited 5 minutes and flipped over the micro plate, then gently tapped on the paper towel, made sure to avoid splashing sample back into wells. Used multi channel pipette to load 150μl of ELISA wash buffer to each well, then flipped over the micro plate and gently tapped on the paper towel. Repeated 1 more time. Used new tip to pipette 50μl of 1°Ab to each well and waited 5 minutes, then flipped over on the paper towel let the liquid out. Used the same step as above to wash 2 times with 150µl of wash buffer. With a new tip to load 50µl of 2°Ab to each well and waited 5 minutes, then poured out and gently tapped on paper towel. Used the same step as above to wash 3 times with 150µl of wash buffer, and then used a new tip to loaded 50µl of enzyme substrate to each well. Viewed the results for the presence of the antigen.ELISA stands for enzyme-linked immunosorbent assay, also known as an enzyme assay, is used to test for many diseases by detecting either the antibody or the antigen. ELISA is a powerful diagnostic tool in human medicine, disease detection, animal, plants, detecting illegal drug used, and testing indoor air quality and food safety. A sample contains many proteins and may or may not contain the antigen, the two unknown antibody will bind to the antigen if presence in a sample, the last was to evaluate the assay results after enzyme substrate added to the wells. Positive and negative controls are used to compare to the test well. The positive control has a strong color change; the negative control has little or no color change.
Materials: Micro plate, multi channel pipette, single channel pipette, micropipette tips, tips disposable cup, ELISA wash buffer, paper towels, reagent reservoir, positive control, negative control, primary antibody (1°Ab), secondary antibody (2°AB), enzyme substrate, unknown sample A and sample B.
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