ELISA Human - Estrogen receptor (ESRs)

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 2 matching solutions for this experiment

Human Total ER alpha/NR3A1 DuoSet IC ELISA

R&D Systems

Upstream tips	Protocol tips	Downstream 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.	-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
-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.

Manufacturer protocol Publication protocol 1 Relevant paper

TransAM® ER Transcription Factor ELISA Kits

Active Motif

Upstream tips	Protocol tips	Downstream tips
	-Mix gently avoid foaming during the dilution of Antibody Binding Buffer. -The developing solution should be warmed to room temperature at least for 1 hour before use. - Keep away from light as the developing solution is light sensitive.	-Discard diluted antibody buffer after use. -After use, discard remaining developing solution.

Protocol tips
-Mix gently avoid foaming during the dilution of Antibody Binding Buffer. -The developing solution should be warmed to room temperature at least for 1 hour before use. - Keep away from light as the developing solution is light sensitive.
Downstream tips
-Discard diluted antibody buffer after use. -After use, discard remaining developing solution.

Manufacturer protocol Publication protocol 1 Relevant paper

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