Human VEGF ELISA Kit (ab100662)

Protocol tips

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.

Publication protocol

The plasma concentrations of ICAM-1, PAI-1, VEGF, and sTM were measured using ELISA kits (Abcam, Cambridge, United Kingdom). SimpleStep ELISA kits were used for ICAM-1, PAI-1, and sTM, whereas the standard kit was used for VEGF. All samples were run in duplicates. Samples with an individual coefficient of variability (CV) greater than 25% were reanalyzed.

All the ELISA tests were performed according to the manufacturer’s instructions with the exception of the washing steps that were performed with an automated washer (Wellwash; ThermoFisher Scientific, Waltham, MA) using a mixture of Tween 20 and phosphate-buffered saline (0.05% vol/vol). The plates were read at 450 nm using an automated microplate reader (Multiskan; ThermoFisher Scientific). The standard curves were constructed using a 4-parameter logistic regression fitting equation (online software: elisaanalysis.com). The mean intra-assay and inter-assay CV for ICAM-1, PAI-1, and VEGF was <10% and <15%, respectively.

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Manufacturer protocol

Download the product protocol from Abcam for Human VEGF ELISA Kit (ab100662) below.

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