Prospective biomarkers selected from the Luminex screen were validated by ELISA. Human SDF-1α ELISA kit (catalog#ELH-SDF-1α), Human IL-8 ELISA kit (catalog #ELH-IL-8), Human IL-1ra ELISA kit (catalog# ELH-1L-1ra), and Human IL-1α ELISA kit (catalog#ELH-IL-1α) were purchased from Ray Biotech, GA and used following manufacturer protocol. Urine samples were diluted 1 in 20 for IL-8 and IL-1ra and 1 in 2 for IL1-α and SDF-1α. Levels of urine creatinine were assayed using the Creatinine Parameter Assay Kit (catalog#KEG005, R&D systems). The absolute levels of urine protein biomarkers were determined using standard curves run on each ELISA plate, and normalized by urine creatinine concentration.
Human sdf 1α elisa kit
Manufactured by RayBiotech
The Human SDF-1α ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) designed for the quantitative measurement of human SDF-1α (Stromal Cell-Derived Factor 1 alpha) levels in biological samples. The kit employs specific antibodies coated on a 96-well plate to capture and detect the target analyte.
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2 protocols using human sdf 1α elisa kit
1
Validating Urine Biomarkers by ELISA
2
Comparative ELISA Evaluation of SDF-1 Kits
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Two different lot numbers of CXCL12/SDF-1 DuoSet were purchased from RnDsystems (catalog N° DY460). The kit consists of a mouse anti-human/mouse SDF1α capture antibody, a biotinylated goat anti-human/mouse SDF1α detection antibody, and a Streptavidin-horseradish-peroxidase-(HRP) conjugate.
Experiments were repeated with two other ELISA kits: the human SDF1α mini ELISA Development Kit (Peprotech; catalog N° 900-M92) which consists of an anti-human SDF1α capture antibody, a biotinylated anti-human SDF1α detection antibody and an avidin-HRP conjugate and the human SDF1α ELISA Kit (Raybiotech; catalog N° ELH-SDF1a) which includes a plate precoated with capture antibody, a biotinylated anti-human SDF1α detection antibody, and a streptavidin-HRP conjugate.
The analyses were performed in Nunc Maxisorp 96-well plates for the RnD and Peprotech kit and in the supplied plate for the Raybiotech kit, according to manufacturer’s instructions. The readout was performed in a Tecan Infinite M200 microtiter plate reader.
Experiments were repeated with two other ELISA kits: the human SDF1α mini ELISA Development Kit (Peprotech; catalog N° 900-M92) which consists of an anti-human SDF1α capture antibody, a biotinylated anti-human SDF1α detection antibody and an avidin-HRP conjugate and the human SDF1α ELISA Kit (Raybiotech; catalog N° ELH-SDF1a) which includes a plate precoated with capture antibody, a biotinylated anti-human SDF1α detection antibody, and a streptavidin-HRP conjugate.
The analyses were performed in Nunc Maxisorp 96-well plates for the RnD and Peprotech kit and in the supplied plate for the Raybiotech kit, according to manufacturer’s instructions. The readout was performed in a Tecan Infinite M200 microtiter plate reader.
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