Mouse STC2 ELISA Kit

Cat No.: 32C040                                 ★Download Datasheet★

Introduction

Stanniocalcin 2 (STC2) is a glycosylated, disulfide-linked, homodimeric hormone initially identified in the corpus of stannous--a small endocrine organ of bony fish. In mammals, STC2 is widely expressed in different tissues as well as in tumor cells. STC2 has been
implicated in diverse biological processes, including calcium and phosphate regulation, cytoprotection, cell development, and
angiogenesis , and is a potential prognostic marker for variety of cancers . STC2 overexpression in MC3T3 cells facilitated
osteoblast differentiation and mineralization through regulation of ERK1/2 phosphorylation, suggesting its involvement in bone
metabolism. Additionally, STC2 has been shown to attenuate ovarian progesterone biosynthesis via PKA pathway, accompanied with the inhibition of follicle-stimulating hormone (FSH)-induced Cyp1a1 and 3β-hydroxysteroid dehydrogenase expression. Furthermore, STC2 is a potential metabolic hormone involved in regulation of glucose homeostasis. In both brown adipose tissue and liver of mice, STC2 expression is induced by fasting, and treatment with recombinant STC2 increases glycogen levels in the fasting state, and
exerts opposite effects on gluconeogenesis in rat hepatocytes in the fasting and fed states.

Principle of the Assay

This assay is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA). The microtiter plate is precoated with affinity
purified polyclonal antibody against mouse STC2. Standards and samples are pipetted into the wells and any mouse STC2 present
is bound by the immobilized antibody. After washing away any unbound substances, a biotin-labelled polyclonal antibody against
mouse STC2 is added to the wells. After washing step to remove any unbound reagents, streptavidin-horseradish peroxidase
(STP-HRP) conjugate is added. After the last wash step, an HRP substrate solution 3,3′,5,5′-Tetramethylbenzidine (TMB) is added,
and color develops in proportion to the amount of mouse STC2 bound initially. Color reaction is stopped by 2M H2SO4 and the
optical density of the wells are determined using a microtiter plate reader at 450nm. Since the increases in absorbance are directly
proportional to the amount of captured mouse STC2, the unknown sample concentration can be calculated from the standard curve
included in each assay.

Assay Performance

TYPICAL STANDARD CURVE

The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample assay.

Assay Characteristics

A. Sensitivity

The lowest level of mouse STC2 that can be measured by this assay is 0.156 ng/mL.

B. Precision

Intra-assay Precision (Precision within an assay)

Two samples of known concentration were tested 8 times on one plate.

Inter-assay Precision (Precision between assays)

Two samples of known concentration were tested in 8 separate assays.

C. Linearity and Recovery

To assess the linearity of the assay, samples spiked with high concentrations of mouse STC2 were serially diluted with the 1×Assay buffer to produce samples with values within the dynamic range of the assay.

Sample1

D. Spike and recovery

Serum samples were assayed by adding 90 μL of sample and 10 μL of spike stock solution calculated to yield the intended 0, 0.625, 2.5 or 10 ng/mL spike concentration.