ELISA Human - GM-CSF

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

Human GM-CSF DuoSet 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

GM-CSF Human ELISA Kit

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
-All samples and reagents must be at room temperature (20-25°C) before use in the ELISA. -Do not use water baths to thaw samples. Thaw at room temperature.	-When preparing standard curve and sample dilution in culture medium, use the same medium used to culture the cells. -Once reagents have been added to the plate, take care NOT to let plate DRY at any time during the assay. -If the TMB substrate solution is blue before use, DO NOT USE IT. -Prepare Streptavidin-HRP Solution immediately before use.

Upstream tips
-All samples and reagents must be at room temperature (20-25°C) before use in the ELISA. -Do not use water baths to thaw samples. Thaw at room temperature.
Protocol tips
-When preparing standard curve and sample dilution in culture medium, use the same medium used to culture the cells. -Once reagents have been added to the plate, take care NOT to let plate DRY at any time during the assay. -If the TMB substrate solution is blue before use, DO NOT USE IT. -Prepare Streptavidin-HRP Solution immediately before use.

Manufacturer protocol Publication protocol 1 Relevant paper

Multi-Analyte ELISArray Kits

Qiagen

Upstream tips	Protocol tips	Downstream tips
Briefly centrifuge any 1.5 ml tube in the kit before opening it in order to collect its contents at the bottom of the tube.	-Visually inspect the Wash Buffer Concentrate to ensure that all components are in solution. If any precipitates are visible, briefly shake the bottle to suspend any precipitate. -Avoid prolonged exposure of the Development Solution to light or contact with water, air, or extreme temperature.

Upstream tips
Briefly centrifuge any 1.5 ml tube in the kit before opening it in order to collect its contents at the bottom of the tube.
Protocol tips
-Visually inspect the Wash Buffer Concentrate to ensure that all components are in solution. If any precipitates are visible, briefly shake the bottle to suspend any precipitate. -Avoid prolonged exposure of the Development Solution to light or contact with water, air, or extreme temperature.

Manufacturer protocol Publication protocol 1 Relevant paper

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