Principle of the Assay
This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- Aβ42
antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- Aβ42 antibody
was used as detection antibodies. The standards, test samples ａnd biotin conjugated detectionantibody were added to the wells subsequently, ａnd washed with wash buffer. HRP-
Streptavidin was added ａnd unbound conjugates were washed away with wash buffer. TMB
substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to
produce a blue color product that changed into yellow after adding acidic stop solution. The
density of yellow is proportional to the Aβ42 amount of sample captured in plate. Read the
O.D. absorbance at 450nm in a microplate reader, ａnd then the concentration of Aβ42 can be
calculated.
Precautions
1. To inspect the validity of experiment operation ａnd the appropriateness of sample dilution
proportion, pilot experiment using standards ａnd a small number of samples is
recommended.
2. After opening ａnd before using, keep plate dry.
3. Before using the Kit, spin tubes ａnd bring down all components to the bottom of tubes.
4. Storage TMB reagents avoid light.
5. Washing process is very important, not fully wash easily cause a false positive.
6. Duplicate well assay is recommended for both standard ａnd sample testing.
7. Don’t let Micro plate dry at the assay, for dry plate will inactivate active components on
plate.
8. Don’t reuse tips ａnd tubes to avoid cross contamination.
9. Avoid using the reagents from different batches together.
Material Required but Not Supplied
1. Microplate reader (wavelength: 450nm)
2. 37°C incubator
3. Automated plate washer
4. Precision single ａnd multi-channel pipette ａnd disposable tips
5. Clean tubes ａnd Eppendorf tubes
6. Deionized or distilled water
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