Human Insulin-like growth factor II (IGF-II) ELISA Kit

Sample

Serum and plasma

Species Reactivity

Human

Detection Method

sELISA

Intended Use

Human IGF-II ELISA Kit is suited for measuring IGF-II, human Insulin-like Growth Factor-II, in human serum and plasma for diagnostic and scientific purposes.

Contents of Kit

1. Microtiter plate: ready for use, coated with mouse-anti-hIGF-II-antibody. Wells are separately breakable. (8x12) wells
2. Calibrators (CAL A-E): lyophilized, (recombinant human hIGF-II), concentrations are given on vial labels and on quality certificate. 5 x 500 μL
3. Control 1 (CTR1): lyophilized, (human serum), concentration is given on quality certificate. 1 x 250 μL
4. Control 2 (CTR2): lyophilized, (human serum), concentration is given on quality certificate. 1 x 250 μL
5. Antibody Conjugate (DET): ready for use, contains goat biotinylated anti-hIGF-II antibody. 1 x 6 mL
6. Enzyme Conjugate (EC): ready for use, contains HRP (Horseradish-Peroxidase)-labeled Streptavidin. 1 x 12 mL
7. Sample Buffer (SB): ready for use. Please shake before use! 1 x 125 mL
8. Washing Buffer (WB): 20-fold concentrated solution. 1 x 50 mL
9. Substrate (S): ready for use, horseradish-peroxidase-(HRP) substrate, stabilised Tetramethylbencidine. 1 x 12 mL
10. Stopping Solution (STP): ready for use, 0.2 M sulfuric acid. 1 x 12 mL
11. Sealing Tape: for covering the microtiter plate. 2

Storage

Store the kit at 2-8°C after receipt until its expiry date. The lyophilized reagents should be stored at –20 °C after reconstitution. Avoid repeated thawing and freezing.
The shelf life of the components after initial opening is warranted for 4 weeks, store the unused strips and microtiter wells airtight together with the desiccant at 2-8°C in the clip-lock bag, use in the frame provided. The reconstituted components Calibrators A-E and Controls CTR1 and CTR2 must be stored at –20°C (max. 4 weeks). For further use, thaw quickly but gently (avoid temperature increase above room temperature and avoid excessive vortexing). Up to 3 of the freeze-thaw cycles did not influence the assay. The 1:20 diluted Washing Buffer WP is 4 weeks stable at 2-8°C

Precision

The Inter- and Intra-Assay variation coefficients are in mean <10%.
Intra-Assay Variation

Inter-Assay Variation

Sensitivity

Sensitivity was assessed by measuring the blank and calculating the theoretical concentration of the 2-fold standard deviation of the blank. The mean analytical sensitivity of the Human IGF-II ELISA Kit is 0.06 ng/mL (Range 0.01 - 0.223 ng/mL, n = 7)

General Description

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). Beside the high affinity Insulin-like Growth Factor Binding Proteins 1-6, IGFs are also bound 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 IGFII 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 notreproducible recoveries.
Human IGF-II ELISA Kit 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.
Indication
Scientific studies in the context of neonatal hypertrophy or hypotrophy (IGF-II is a fetal growth factor) and malignancy (IGF-II is an oncogenic growth factor). IGF-II appears to be suitable for differential diagnosis in various malignant diseases. For example, IGF-II can be used to differentiate between adrenocortical tumors and adenomas. In prostate cancer, too, tumor staging and the differentiation between carcinoma and hyperplasia can be improved by measuring IGF-II in serum. Recent results from neurology show that the IGF system is also important in the development of Alzheimer's and Parkinson's disease.

Standard Curve

The following data is for demonstration only and cannot be used in place of data generation at the time of assay.

The exemplary shown standard curve in Figure below cannot be used for calculation of your test results. You have to establish a calibration curve for each test you conduct!