Ghrelin Unacylated Rat ELISA

Research topic

Animal studies, Energy metabolism and body weight regulation

Features

• The kit measures plasma unacylated ghrelin.
• Quality controls are rat-plasma based.

Storage/Shipping

Store the kit at –20°C. Under these conditions, the kit is stable until expiration date (see label on the box).

Summary

Ghrelin, an endogenous ligand for the growth hormone secretagogue receptor, is synthesized principally in the stomach. It stimulates food intake and transduces signals to hypothalamic regulatory nuclei that controls energy homeostasis. The peptide consists of 28 amino acids, with a n-octanoylation of the serine-3 residue, which is necessary for the biological activity mentioned below. Ghrelin is present in the peripheral circulation under two forms: acylated and unacylated.

Assay format

Sandwich ELISA, AChE-labelled antibody

Sample requirements

10 µl/well

Applications

Plasma-EDTA

Calibration Curve

Calibration range

1.96 – 250 pg/ml

Limit of detection

The Limit of Determination, calculated as the concentration of unacylated ghrelin corresponding to the NSB average (n = 8) plus three standard deviations is 0.6 pg/ml and 0.8 pg/ml for long and short first incubation period, respectively.

Limit of quantification

Due to the minimal plasma dilution needed (1:10), the limits of quantification in the samples are 6 pg/ml (20h at +4°C) and 8 pg/ml (3h at RT), respectively.

Intra-assay

CV = 8.2 % (20h at +4°C)
CV = 9.2 % (3h at RT)

Inter-assay

CV = 8.6 % (20h at +4°C)
CV = 9.5 % (3h at RT)

Spiking Recovery

101.5 % (20h at +4°C)
99.8 % (3h at RT)

Dilution Linearity

98.9 %