hGH Sensitive ELISA Assay Kit

$520.00

The Eagle Biosciences hGH Sensitive ELISA is an enzyme immunoassay for the quantitative in-vitro diagnostic measurement of levels of human growth hormone in serum and plasma. Human growth hormone measurements are used in the diagnosis and treatment of disorders involving the anterior lobe of the pituitary gland. The Eagle Biosciences hGH Sensitive ELISA Assay kit is for Research Use Only. 

SKU: E022 Categories: , ,

hGH Sensitive ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 12 pg/ml
Incubation Time: 2.75 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL

Manufactured by Mediagnost

Assay Notes:

The 2nd International Standard for hGH, NIBSC Code 98/574 (6), was used as standard material. The International Standard was defined in an international study in the year 2001 with 3 International units per mg Protein (3 IU/mg). The exclusive application of this standard material is recommended in line with the current standardisation efforts for hGH Immunoassays.

Additional Information

Assay Background

The endocrine system of human Growth Hormone (hGH), also named Somatropin, is characterized by an extreme complexity. hGH is the product of the GH-1 gene located on chromosome 17 and expressed in pituitary cells. 80% of the gene expression results in a non-glycolsylated 22 kDa protein consisting of 191 amino acids. The other 20% of gene expression results in a variant form of 20 kDa by alternative splicing. Additionally, several smaller variants can be found in circulation as well as translational modified proteins and different degrees of protein aggregation. Further on, bioactivity of Growth Hormone is regulated by a specific binding protein (GHBP) formed by the extracellular part of the cellular transmembrane GH-receptor. These modifications allow a tight control of the half-life period hGH and of its bioactivity.

Not only synthesis and posttranslational modification but also secretion of hGH is tightly regulated. Spontaneous pulsatile secretion takes place with a single pulse every three hours and a maximal secretion during night’s sleep. Several different attractions as physiologic stress or cross-reactivity and amino acids result in additional hGH secretion, induced by the hypothalamic hormones Somatostatin and GH-Releasing Hormone (GHRH). Age, sexual steroids, nutritional status, illness and emotions influence the amount of secreted hGH. Because of the multitude of influencing factors the normal quantitative secretion cross-reactivity.

Physiological functions of hGH are also manifold. These functions are partially exerted by Insulin-like Growth Factors (IGFs). In children and adolescent, the hGH system is the main regulator of growth. If the hGH system fails totally, human growth will end at 120 cm. Besides regulation of growth hGH exerts an anabolic effect on muscle and connective tissue as wells as on bone and different other organs (heart, intestine). Further hGH was proved to have a lipolytic effect.

Growth Hormone pathology is characterized by extreme high or extreme low hGH secretion. During childhood, it is the Growth Hormone deficiency congenital or acquired, which leads to microsomia. For diagnosis of Growth Hormone deficiency an hGH stimulation test has to be done or the spontaneous excretion must be investigated. The therapy consists of substitution of endogenous Growth Hormone by recombinant hGH resulting in normalization of growth. In adulthood hGH deficiency is mostly caused by pituitary adenoma (and their surgical excision). hGH deficiency shows typical disease pattern, equivalent to advanced aging (adipositas, muscle dystrophy, arteriosclerosis, osteoporosis, adynamia). Substitutional therapy is a well-known, approved and efficient therapy of severe Growth Hormone deficiency in adulthood. Therapeutic success is directly as well as indirectly proved by measurement of IGF in serum.

Excessive hGH secretion, mostly caused by pituitary adenoma, results in childhood in gigantism, in adulthood in acromegaly, leading to enlarged extremities, diabetes, heart insufficiency and tumor growth. Surgical excision of the adenoma is the therapy of choice. If tumor excision is not possible or incomplete, a medicinal therapy with somatostatin preparation will be conducted, resulting in inhibition of hGH production. Alternatively, hGH analoga (e.g. Pegvisomant) are used to block the hGH receptor and thereby inhibit action of endogenous hGH. Determination of human Growth Hormone (hGH, Somatropin) is done for diagnosis of Growth Hormone deficiency or Growth Hormone excess (arcomegaly). During medicinal and/or after surgical therapy of arcomegaly Growth Hormone (and IGF-I) measurement is used for therapy control.

Assay Principle

The Eagle Biosciences hGH Sensitive ELISA is a so-called sandwich-assay. It utilizes a specific, high affinity polyclonal rabbit antiserum coated on the wells of a microtiter plate. The hGH in the samples binds quantitatively to the immobilized antiserum. In the following step, the biotinylated antiserum in turn binds hGH. After washing, a streptavidin-peroxidase-enzyme conjugate will be added, which will bind highly specific to the biotin of the antiserum and will catalyze the substrate to change the color quantitatively depending on the hGH level of the sample.

Specificity

The only human protein with significant sequence similarities to growth hormone is prolactin. Testing a 200 ng/mL prolactin solution in this assay, no cross-reactivity was detected.

Further, Pegvisomant (trade name Somavert) a growth hormone analogue and pharmaceutical used in acromegaly therapy has been tested for cross-reactivity in different concentrations. Here no significant influence of Somavert was detected (Table 1). But in Growth Horm IGF Res 18 (2008) 526-532 the authors added 100 mg/L pegvisomant to hGH enriched samples containing 2.6 and 10.1 µg/L hGH and measurement results were 154% and 108% of the expected values respectively.

Manual

Product Manual