Mouse/Rat IGFBP-3 (insulin-like growth factor binding protein 3) ELISA Kit

Sample

Serum, plasma and cell culture medium

Species Reactivity

Mouse and rat

Detection Method

sELISA

Intended Use

This enzyme immunoassay kit is suited for measuring IGFBP-3 in mouse and rat serum and plasma and in cell culture medium.

Contents of Kit

1. Microtiter plate: ready for use, coated with goat anti-mouse-IGFBP-3 antibody. Wells are separately breakable. (8x12) wells
2. Standards (A-G): lyophilized, (native mouse IGFBP-3), concentrations are given on vial labels and on quality certificate. 7 x 750 μL
3. Control Serum 1 (KS1): lyophilized, (mouse serum), concentration is given on quality certificate. 1 x 250 μL
4. Control Serum 2 (KS2): lyophilized, (mouse serum), concentration is given on quality certificate. 1 x 250 μL
5. Antibody Conjugate (AK): ready for use, goat-anti-mouse-IGFBP-3 antibody biotinylated. 1 x 12 mL
6. Enzyme Conjugate (EK): contains HRP (Horseradish-Peroxidase)-labeled Streptavidin. 1 x 12 mL
7. Dilution Buffer (VP): ready for use. Please shake before use! 1 x 125 mL
8. Washing Buffer (WP): 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 (SL): 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 standards A-G and Control Sera KS1 and KS2 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 were on average ≤10%.
Intra-Assay-Variation (n=13)

Inter-Assay-Variation (n=26 or 15)

Sensitivity

The analytical Sensitivity was assessed by 21-fold determination of the blank and calculating the theoretical concentration of the blank +2SD. The analytical sensitivity of the Mouse/Rat IGFBP-3 ELISA Kit is 0.09 ng/mL.

General Description

Growth Hormone, Insulin-like Growth Factors and their binding proteins build up an endocrine system regulating not only longitudinal growth in humans but also influencing a broad variety of other physiological and pathophysiological processes like energy metabolism or tumor growth. Most effects of Growth Hormone (GH) are exerted by Insulin-like Growth Factors (IGF) mainly produced by the liver but also locally by specific tissues. The effects of IGF are also regulated, specific binding proteins (IGFBP 1-7) regulate bioavailability of IGF. After proteolytic cleavage of the binding proteins IGF is set free and able to bind to its receptor. The autophosphorylation of this thyrosine kinase receptor activates intra cellular signalling cascades. Some of these IGFBPs not only regulate the availability of IGF but also exert IGF-independent effects on cell physiology.
IGFBP-3 is the most abundant IGFBP in circulation and therefore of special relevance in regulation of IGF effects. This is reflected by the indicative value of serum IGFBP-3 concentration in diagnostics of growth disturbances. Regulation is effected e.g. through nourishing situation; Different diets for example affect the IGFBP-3 concentration (Bielohuby et al, 2010). IGFBP-3 has also been shown to be able to induce apoptosis, promote tumor growth and inhibit cellular migration and metastasis dependent on tissue and tumor stage. Mouse / rat models for in vivo experiments are often used for studies of IGF-dependent and independent effects of IGFBP-3, particularly in the field of tumor research.

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 calibration 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!