Human NGAL DuoSet ELISA kit (R&D, catalog# DY1757, lot# P195696) was employed in the study. Specifically, 100 μl of serial diluted standard samples (PART#842273) as well as urinary samples from patients and self-described healthy volunteers (10-fold dilution using reagent diluent) were added into different wells and the plate was incubated at room temperature for 2 hours. The plate was washed subsequently and incubated with biotinylated detection antibodies (PART#844865, 25 ng/ml in reagent diluent) for 2 hours, washed again with PBST, and incubated with HRP-labeled streptavidin (PART#893975, 40-fold dilution using reagent diluent) for 20 mins. Human NGAL p-FLISA was performed by adopting a similar procedure as the ELISA with significantly shortened assay time. Specifically, the incubation time for standards/samples, biotinylated detection antibodies, 800CW-labeled streptavidin (LI-COR P/N 926-32230, 50 ng/ml), and plasmonic-fluor-800CW (extinction~1) was set to 5 minutes. The study was approved by Washington University IRB 201601082 “Nanotech Biomarkers for Renal Cancer Intervention: Clinical Validation and Utility” and IRB 201202051 “Urine Proteome of Surgical Patients and Healthy Volunteers”. Informed consent was obtained from the participants.
Human ngal duoset elisa kit
Manufactured by R&D Systems
The Human NGAL DuoSet ELISA kit is a laboratory assay designed to quantitatively measure the levels of the protein NGAL (Neutrophil Gelatinase-Associated Lipocalin) in human samples. The kit provides the necessary components to perform a sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of NGAL.
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2 protocols using human ngal duoset elisa kit
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Rapid NGAL Detection in Urine Samples
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Rapid NGAL Detection in Urine Samples
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Human NGAL DuoSet ELISA kit (R&D, catalog# DY1757, lot# P195696) was employed in the study. Specifically, 100 μl of serial diluted standard samples (PART#842273) as well as urinary samples from patients and self-described healthy volunteers (10-fold dilution using reagent diluent) were added into different wells and the plate was incubated at room temperature for 2 hours. The plate was washed subsequently and incubated with biotinylated detection antibodies (PART#844865, 25 ng/ml in reagent diluent) for 2 hours, washed again with PBST, and incubated with HRP-labeled streptavidin (PART#893975, 40-fold dilution using reagent diluent) for 20 mins. Human NGAL p-FLISA was performed by adopting a similar procedure as the ELISA with significantly shortened assay time. Specifically, the incubation time for standards/samples, biotinylated detection antibodies, 800CW-labeled streptavidin (LI-COR P/N 926-32230, 50 ng/ml), and plasmonic-fluor-800CW (extinction~1) was set to 5 minutes. The study was approved by Washington University IRB 201601082 “Nanotech Biomarkers for Renal Cancer Intervention: Clinical Validation and Utility” and IRB 201202051 “Urine Proteome of Surgical Patients and Healthy Volunteers”. Informed consent was obtained from the participants.
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