IGF-II ELISA Assay Kit

$620.00

The IGF-II ELISA Assay Kit measures IGF-II in human serum, plasma, urine, saliva as well as cell culture supernatants. The IGF II ELISA Assay is for research use only and not for use in diagnostic procedures.

IGF-II ELISA Assay Kit

The IGF-II ELISA Assay Kit is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.06 ng/ml
Ranage: 0.06-3636 ng/ml
Incubation Time: 3 hours
Sample Type: Serum, Plasma, urine, saliva and cell culture medium
Sample Size: 10 µL

Product is manufactured by Mediagnost


Assay Notes:

The standards of the ELISA E30 are human IGF-II in concentrations 0.45; 1.5; 3; 5.63 and 9 ng/ml. The assay range covers the range to 2400 ng/ml. By varying the sample dilution this can be adapted to the special individual requirements. The Eagle Biosciences IGF-II ELISA Assay Kit is calibrated against the International Standard: WHO NIBSC 96/538.


Assay Background

The insulin-like growth factors (IGF)-I and –II play a pivotal role in the regulation of proliferation and differentiation of several tissue types. IGF-I also called Somatomedin C has a molecular weight of 7.469 kDa. Its expression is mainly regulated by Growth Hormone and nutrition. But several hormones and peptide factors are known to influence IGF-II synthesis in different tissues. Bioavailability of the IGFs is regulated by specific binding proteins (IGFBP). Besides the high-affinity Insulin-like Growth Factor Binding Proteins 1-6, IGFs are also bound to be IGFBP-related Proteins. These binding proteins bind IGF-I and IGF-II with the same affinity or prefer IGF-II. Direct measurement of IGFs in serum samples without pretreatment results in false values because of the extremely slow dissociation of the IGF/IGFBP complexes during the assay incubation only a part of the IGF-II in the specimen can bind to the antibodies and be detected.

Therefore, various techniques were applied to physically separate IGF-II from its binding proteins before measurement, including (a) size exclusion chromatography under acidic conditions, (b) solid-phase extraction and (c) acid-ethanol extraction. These techniques, however, are either inconvenient or time-consuming or give incomplete and not-reproducible recoveries.

This assay is easy, fast and results do not depend on the binding protein concentration of the sample. It is based on the high specificity of the employed antibodies for IGF-II. There is virtually no cross-reactivity with IGF-I. This allows the separation of IGF-II from the binding proteins by acidification and blocking of the free binding proteins with IGF-I. Thus, the endogenous IGF-II is free in solution.


Related Products

IGF-I ELISA Assay Kit
Human IGFBP-1 ELISA Assay
IGFBP-6 ELISA Assay Kit

Additional Information

Assay Principle


The Eagle Biosciences ELISA for IGF-II is a so-called Sandwich-Assay. It utilizes two specific and high-affinity antibodies for this protein. The first antibody, immobilized on the microtiter plate, and the added second biotinylated antibody are binding the IGF-II in the sample. The Streptavidin-Peroxidase Enzyme Conjugate subsequently binds to the complex. In the closing substrate reaction, the turn of the colour will be high specific catalysed, quantitatively depending on the IGF-II-level of the samples.

IGF-II-IGFBP complex is dissociated by dilution in an acidic buffer. IGFBPs are blocked by IGF-I excess, thus allowing the measurement of free IGF-II. With this method, the IGFBPs are not removed, but their function and therefore their interference in the assay is neutralized.  Due to the low cross-reactivity of the IGF-II antibody with IGF-I, excess IGF-I does not disturb the interaction of the first antibody with IGF-II.

Specificity


Because of the high specificity of the used antibodies the endogenous and structurally very similar IGF-I existing in variable concentration in the samples does not affect the correctness of the IGF-II determination up to 1000 ng/ml.

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