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AGRP (83-132) Amide Fluorescent EIA Kit Maximize

AGRP (83-132) Amide Fluorescent EIA Kit

Human AGRP (83-132) Amide Fluorescent EIA Kit (FEK-003-53) designed for quantitatively detecting amounts of human agouti-related protein (83-132) amide (AGRP-NH2).

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FEK-003-53

96 wells

£ 480.00

Human AGRP Amide Fluorescent EIA Kit

Specificity : Human
Sensitivity : 11.2 pg/ml
Linear Range: 11.2-108 pg/ml
Range: 0-10000 pg/ml
Size: 96 tests

Reagents Supplied:
- 96 well immunoplate (1 plate)
- 20x assay buffer concentrate (50ml)
- Standard peptide (1 vial).
- Primary antibody (rabbit anti-peptide IgG) (1 vial).
- Positive control (1 vial).
- Acetate plate sealer (APS), (3 pieces)
- Streptavidin-horseradish peroxidase (SA-HRP) (30μl)
- Substrate solution (12ml)
- Biotinylated peptide (1 vial).
- Stable peroxide solution (1.5ml)
- Stop solution (12ml)
- General protocol (1 book).
- Assay diagram (1 sheet).

Intended Use:
Human AGRP Amide Fluorescent EIA kit is a quantitative procedure to analyse concentrations of human agouti-related protein (83-132) amide using serum, any biological fluid samples, plasma, urine or CSF samples.

Assay Principle:
1. Human AGRP amide fluorescent EIA has been pre-coated with secondary antibody and also the non-specific binding sites have been blocked.
2. Secondary antibody is able to bind with the Fc fragment of the primary antibody, the Fab segment is competitively bound by both biotinylated peptides and the peptide standard or the targeted peptide in samples.
3. Biotinylated peptide can then interact with the SA-HRP, which then catalyzes the substrate solution.
4. Fluorescence intensity is directly proportional to the amount of biotinylated peptide-SA-HRP complex but inversely proportional to the amount of the peptide in standard solutions or samples.
5. Unknown concentration in samples can then be determined by using this human AGRP amide fluorescent EIA kit to extrapolation from a standard curve.

Cross Reactivity:
AGRP (83-132)-NH2 (Human) 100%.
AGRP Form C-Amide (Human) 100%.
AGRP (82-131)-NH2 (Mouse) 65%.
Orexin B (Human) 0%.
Orexin A (Mouse, Rat, Human) 0%.
Leptin (Human) 0%.
MCH (Rat, Human, Mouse) 0%.
alpha MSH 0%.
CGRP (Rat, Human) 0%
NPY 0%.GGRP (Human, Rat) 0%.

Background:
Agouti-related peptide (AGRP) is paracrine signalling molecule which is composed of 112 amino acids. Understanding the function AGRP plays in weight gain could assist in help reduce obesity. Overexpression in transgenic mice results in obesity and hyperphagia. It found to be mainly expressed in the following: hypothalamus, adrenal gland, subthalamic nucleus. It increases appetite and decreases metabolism and energy expenditure. AGRP has been shown to be an inverse agonist of melanocortin receptors, specifically, MC3-R and MC4-R.

References
1. Cell Metab. (2011) 14 (2): 173-83. Autophagy in hypothalamic AgRP neurons regulates food intake and energy balance. Kaushik S, et al.
2. J Neurol. (2013) 260 (2): 475-83. Role of intestinal peptides and the autonomic nervous system in postprandial hypotension in patients with multiple system atrophy. Takeshi Fukushima, et al.
3. Biochem. Biophys. Res. Commun. (2003) 301 (1): 102-7. Agouti-related protein has an inhibitory paracrine role in the rat adrenal gland. Dhillo WS, et al.
4. Neurosci Lett. (2004) 370 (2-3): 108-13. The effects of agouti-related protein gene transfer in vivo by electroporation in mice. Xiang, et al.
5. Genes Dev. (1997) 11 (5): 593-602. Hypothalamic expression of ART, a novel gene related to agouti, is up-regulated in obese and diabetic mutant mice. Shutter JR, et al.

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