Human Nucleobindin-2 PharmaGenie ELISA Kit (SBRS0100)

Nucleobindin-2 (NUCB2) is a calcium-binding EF-hand protein that is post-translationally cleaved into nesfatin-1, an anorexigenic peptide implicated in satiety signaling, energy homeostasis, and glucose metabolism. Altered NUCB2/nesfatin-1 levels have been investigated in research on obesity, type 2 diabetes, polycystic ovary syndrome, and stress-driven appetite suppression, supporting its role as a candidate biomarker in metabolic and endocrine studies.

This Assay Genie PharmaGenie kit is built on a competitive immunoassay architecture with an absorbance-based chromogenic readout. Reconstituted standards are run in a serial dilution alongside unknown samples to construct the calibration curve, and absorbance values are interpolated against this curve to determine NUCB2 concentrations in each well. The protocol uses a biotinylated detection reagent and a streptavidin-HRP conjugate, with a sample volume of 50 µL loaded per well, and absorbance is read on a standard microplate reader after addition of the chromogenic substrate and stop solution.

The assay is validated for cell culture supernatant and serum-derived matrices, supporting both in vitro experiments on adipose, hypothalamic, and pancreatic cell models and analysis of clinical research samples, with cross-reactivity characterized against the full-length NUCB2 precursor and the nesfatin-1 cleavage product. The plate provides 96 sample wells per run, sufficient for full standard curves, internal quality controls, and replicate measurements across multiple treatment conditions, and the assay covers a working dynamic range of approximately 0.1–10 ng/mL with a sensitivity near 0.05 ng/mL.

Storage and handling guidance: refrigerate unopened components, protect the chromogenic substrate from light, and avoid repeated freeze-thaw cycles of reconstituted standards and working solutions. Allow plates and buffers to equilibrate to room temperature before initiating the assay to ensure reproducible well-to-well measurements.