ELISA Human - ICAM-1/CD54

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 ICAM-1/CD54 Allele-specific Quantikine ELISA Kit

R&D Systems

Upstream tips	Protocol tips	Downstream tips
-Bring all reagents to room temperature before use.	-When mixing or reconstituting protein solutions, always avoid foaming. -To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent. -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.

Upstream tips
-Bring all reagents to room temperature before use.
Protocol tips
-When mixing or reconstituting protein solutions, always avoid foaming. -To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent. -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.

Manufacturer protocol Publication protocol 1 Relevant paper

Human ICAM1 ELISA Kit (CD54) (ab100640)

Abcam

Upstream tips	Protocol tips	Downstream tips
-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. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.	-Unused well strips should be returned to the plate packet and stored at 4°C.

Upstream tips
-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. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.
Downstream tips
-Unused well strips should be returned to the plate packet and stored at 4°C.

Manufacturer protocol Publication protocol 1 Relevant paper

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