Human IGFBP-2 (insulin-like growth factor binding protein 2) ELISA Kit

Sample

Serum, plasma or other body fluids

Species Reactivity

Human

Detection Method

sELISA

Intended Use

This enzyme immunoassay kit is suited for measuring IGFBP-2 in human serum, plasma or in other body fluids, for example, cerebrospinal fluid, breast milk, amniotic fluid, saliva, as for scientific purposes. It is also suited to quantitate IGFBP-2 in cell culture media.

Contents of Kit

1. Microtiter plate: ready for use, coated with mouse-anti-hIGFBP-2-antibody. Wells are separately breakable. (8x12) wells
2. Calibrators (CAL A-E): lyophilized, (recombinant human IGFBP-2), concentrations are given on vial labels and on quality certificate. 5 x 750 μL
3. Control 1 (CTR1): lyophilized, (human serum), concentration is given on quality certificate. 1 x 100 μL
4. Control 2 (CTR2): lyophilized, (human serum), concentration is given on quality certificate. 1 x 100 μL
5. Antibody-HRP-Conjugate (DET): ready for use, contains rabbit biotinylated anti-hIGFBP-2 antibody + streptavidin horseradish peroxidase conjugate. 1 x 12 mL
6. Dilution Buffer (DIL): ready for use. 1 x 50 mL
7. Washing Buffer (WB): 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 (STP): 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 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 coefficients of variability are below 10%.

Sensitivity

Sensitivity was assed by measuring the blank and calculating the theoretical concentration of the blank + 2SD. The analytical sensitivity of the Human IGFBP-2 ELISA Kit is 0.2 ng/mL.

General Description

Insulin-like growth factors (IGFs) regulate the proliferation, differentation, apoptose, cell adhesion and metabolism in various tissues and cell types. The IGF-I, which is produced mainly in liver under the influence of growth hormone (GH), regulates as hormone the linear growth of the bones and the process of sexual maturity, while IGF-II is mainly a growth factor of fetal tissue. The biological actions of IGF over the IGF-Type-I receptor are modulated variably through the IGF binding proteins (IGFBP-1 to-6). IGFBP-2 is, after IGFBP-3, the second most frequent IGFBP in the human blood. IGFs, especially tumor typical pro-IGF-forms and hormones regulate the expression of IGFBP-2, GH effect is thereby inhibiting. At cellular level IGFBP-2 seems to stimulate the proliferation and dissemination of solid tumors via an IGF-independent mechanism. IGFBP-2 is a 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 IGFBPserum levels after remission. During the GH-therapy, e.g. in short stature and in GHabuse (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.

Standard Curve

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!