ELISA Human - Adiponectin

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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Found 3 matching solutions for this experiment

Upstream tips
-The entire kit may be stored at -20°C for long term storage before reconstitution.
-Avoid repeated freeze-thaw cycles.
-Equilibrate all reagents to room temperature (18-25°C) prior to use.
-Prepare fresh reagents immediately prior to use. If crystals have formed in the concentrate, mix gently until the crystals have completely
dissolved.
Protocol tips
-Prepare serially diluted standards immediately prior to use.
-Always prepare a fresh set of standards for every use.
-Read plate immediately
after adding STOP solution.
Downstream tips
-Store all reagents as
recommended.
-Any remaining standard should be stored at -20°C after reconstitution and used within 30 days.
Human Adiponectin/Acrp30 DuoSet ELISA

R&D system, Minneapolis, MN, USA

Upstream tips
-Bring all reagents to room temperature before use.
-Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution.
-Working dilutions should be prepared and used immediately.
Protocol tips
-A standard curve should be generated for each set of samples assayed.
-It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum.
-Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.
Upstream tips
-Store the complete kit at 2-8°C.
-All reagents need to be brought to room temperature prior to use.
Protocol tips
-Do not make foam when diluting the sample.
-Substrate Solution should remain colourless until added to the plate. Keep Substrate
Solution protected from light.
-Stop Solution should remain colourless until added to the plate.
Downstream tips
-Opened reagents are stable 3 months when stored at 2-8°C.
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