Mouse/Rat IGFBP-3 ELISA Kit

Sample

Serum, plasma, cell culture medium

Species Reactivity

Mouse; Rat

Detection Method

sELISA

Intended Use

This ELISA 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 (96 wells)
2. Standards (7×750 μL)
3. Control Serum 1 (1×250 μL)
4. Kontrollserum 2 (1×250 μL)
5. Antibody Conjugate (1×12 mL)
6. Enzyme Conjugate EK (1×12 mL)
7. Dilution Buffer (1×125 mL).
8. Washing Buffer WP(1×50 mL)
9. Substrate S (1×12 mL)
10. Stopping Solution SL (1×12 mL)

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.

Performance Characteristics

Quantitative determination of mouse/rat IGFBP-3 without sample pretreatment

Precision

The Inter- and Intra-Assay variation coefficients were on average ≤10%. Exemplary determinations are shown in table 1 and table 2.
Table 1 Inter-Assay-Variation (n=26 or 15)

Table 2 Intra-Assay-Variation (n=13)

VC=Coefficient of Variation

Detection Range

0.078-5 ng/ml

Detection Limit

0.018 ng/ml

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 DEIA4628 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. 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. For this purpose CD offers the product as a reliable and sensitive test system for the determination of IGFBP-3 in mouse and rat samples.

Standard Curve

Reconstitution And Storage

The Standards A-G and Control KS1 and KS2 are reconstituted with the Dilution Buffer VP. It is recommended to keep reconstituted reagents at room temperature for 15 minutes and then to mix them thoroughly but gently (no foam should result) with a Vortex mixer.
After reconstitution dilute the Control Sera KS1 and KS2 with the Dilution Buffer VP in the same ratio (1:505) as the sample. The required volume of Washing Buffer WP is prepared by 1:20 dilution of the provided 20-fold concentrate with Aqua dest.