ELISA Human - BRCA2

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

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Upstream tips
-All the reagents should be kept at 4oC upon receipt.
-Bring all kit components and samples to room temperature (18-25oC) before use.
Protocol tips
-Reconstitute the Standard with 1.0mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam).
-Prepare standard within 15 minutes of performing the assay. If crystals have formed in the Wash Solution concentrate (30×), warm to room temperature and mix gently until the crystals are completely dissolved.
-TMB Substrate is easily contaminated. Please protect it from light.
Downstream tips
-The reconstituted Standards can be used only once.
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