Mouse/Rat IGFBP-2 (insulin-like growth factor binding protein 2) ELISA Kit

Sample

Serum and plasma

Species Reactivity

Mouse and rat

Detection Method

sELISA

Intended Use

This enzyme immunoassay kit is suited for measuring IGFBP-2 in mouse and rat serum for scientific purposes.

Contents of Kit

1. Microtiter plate: ready for use, coated with rabbit-anti-IGFBP-2 antibody. Wells are separately breakable. (8x12) wells
2. Standards (A-G): lyophilized, (native mouse IGFBP-2), concentrations are given on vial labels and on quality certificate. 7 x 1 mL
3. Control Serum (KS): lyophilized, (mouse serum), concentration is given on quality certificate. 1 x 250 μL
4. Antibody Conjugate (AK): 100-fold concentrated, goat-anti-mouse-IGFBP-2 antibody biotinylated. 1 x 120 μL
5. Enzyme Conjugate (EK): 100-fold concentrated, Streptavidin Peroxidase-Conjugate. 1 x 120 μL
6. Dilution Buffer (VP): ready for use. Please shake before use! 1 x 120 mL
7. Washing Buffer (WP): 20-fold concentrated solution. 1 x 50 mL
8. Substrate (S): ready for use, horseradish-peroxidase-(HRP) substrate, stabilised Tetramethylbencidine. 1 x 12 mL
9. Stopping Solution (SL): ready for use, 0.2 M sulfuric acid. 1 x 12 mL
10. 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 serum KS 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

Intra-Assay Variance
Four serum samples where measured in 16 independent assays. On average the coefficient of variation was 5.1% (SD 1.3).

Inter-Assay-Variability
Four serum samples where measured in 16 independent assays. On average the coefficient of variation was 5.1% (SD 1.3).

Sensitivity

Sensitivity was assessed by measuring the blank and calculating the theoretical concentration of the blank +2SD. The analytical sensitivity of Mouse/Rat IGFBP-2 ELISA Kit is 0.01 ng/ml.

General Description

Insulin-like growth factors (IGFs) regulate the proliferation, differentation, apoptosis, cell adhesion and metabolism in various tissues and cell types IGFBP-2 is an unglycosylated polypetide of 31.3 kDa, which forms binary IGF-complexes and shows no circadian rhythm in the circulation. The serum concentration of IGFBP-2 increases in fasting, after major surgery and after trauma, but the increasing of the concentration is most intensive in malignant diseases.
The correlation of the IGFBP-2 level to the degree of progression is a striking feature in various tumor types as is the normalization of the IGFBP-serum levels after remission. During the GH-therapy, e.g. in short stature and in GH-abuse (doping) the IGFBP-2 level decreases. In Trisomy 18 IGFBP-2 in maternal serum is decreased and IGFBP-1 is increased; therefore the ratio IGFBP-2 /IGFBP-1 is a marker for this chromosome abnormality.
Transgenic organisms are a good opportunity to investigate the function of genes or proteins. The mouse or rat model is a well-suited system for investigation of the relevance of IGFBP-2 in physiological and pathological processes. Over expression of the IGFBP-2 gene in mice results in a weight reduction of 30% in spleen and moderately reduced weight in other organs. Effects of IGFBP-2 on the organism can be compensated through the modified expression of other IGF Binding proteins.
Especially in tumor biology the mouse and rat systems enable investigation of the systemic relevance of IGFBP-2. IGFBP-2 influences tumor cells as it induces catalase activity in adrenocortical cells. Furthermore IGFBP-2 interacts with tumor cells via its RGD-amino acid sequence and seems to stimulate cell invasion of glioma cells.

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 standard curve for each test you conduct!