ELISA Human - IGF-I

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 IGF-I/IGF-1 Quantikine ELISA Kit

R&D Systems

Upstream tips	Protocol tips	Downstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.	-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Upstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.
Protocol tips
-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Manufacturer protocol Publication protocol 1 Relevant paper

Human IGF1 ELISA Kit (ab100545)

Abcam

Upstream tips	Protocol tips	Downstream tips
-Prepare serially diluted standards immediately prior to use. Always prepare a fresh set of standards for every use. -Equilibrate all reagents to room temperature (18-25°C) prior to use.	-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Do not vortex the standard during reconstitution, as this will destabilize the protein. -Keep the standard dilutions on ice while during preparation, but the ELISA procedure should be done at room temperature.	-Discard working standard dilutions after use as they do not store well.

Upstream tips
-Prepare serially diluted standards immediately prior to use. Always prepare a fresh set of standards for every use. -Equilibrate all reagents to room temperature (18-25°C) prior to use.
Protocol tips
-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Do not vortex the standard during reconstitution, as this will destabilize the protein. -Keep the standard dilutions on ice while during preparation, but the ELISA procedure should be done at room temperature.
Downstream tips
-Discard working standard dilutions after use as they do not store well.

Manufacturer protocol Publication protocol 1 Relevant paper

Human IGF-1 PicoKine™ ELISA Kit

BosterBio

Upstream tips	Protocol tips	Downstream tips
-Store at 4°C for 6 months, at -20°C for 12 months. -Avoid multiple freeze-thaw cycles. -The standard solutions are best used within 2 hours.	-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment. -Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature. -Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection.

Upstream tips
-Store at 4°C for 6 months, at -20°C for 12 months. -Avoid multiple freeze-thaw cycles. -The standard solutions are best used within 2 hours.
Protocol tips
-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment. -Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature. -Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection.

Manufacturer protocol Publication protocol 1 Relevant paper

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