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Insulin Growth Factor-1 ELISA Kit

GWB-6F09CA

40-056-205032

1x96 well plate
$442.00
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Description

Product Name Insulin Growth Factor-1.
Category Growth Factors.
Sample Type Serum.
Sensitivity 4,9 ng/mL.
Description Enzyme linked immunosorbent assay for the determination of Insulin like Growth Factor -1 in human serum.
# of Samples 1X96 Assays.
Intended Use Immunoenzymetric assay for the in vitro quantitative measurement of human Insulin-like Growth Factor-I (IGF-I) in serum.
Background A. Biological activities
Insulin-like growth factor I (IGF-I) or Somatomedin-C (SM-C) is a basic 70 amino acid single chain polypeptide (MW : 7649 Da) similar to proinsulin (50% sequence homology), and to the other well-characterized member of the somatomedin family : IGF II (67AA, 70 % sequence homology with IGF-I). IGF-I is the most important factor, which mediates the growth promoting actions of growth hormone, a pituitary hormone with highly fluctuating blood levels due to pulsatile release. The blood concentration of IGF-I is more stable due to the binding to carrier proteins. The concentration of the predominant binding protein (MW 53000) as well as the production of IGF-I, are regulated by growth hormone. IGF-I is produced by the liver, and other tissues, and it has endocrine, paracrine and autocrine activities. It stimulates growth and regulates differentiation of various tissues, displays insulin-like activities and promotes cartilage growth. Although GH is the most important factor controlling IGF-I secretion and concentration, other factors are also determinant: the age (with a peak at adolescence), the sex, the nutritional status, and other hormones (oestrogen, thyroxin, prolactin,..). Specific trophic stimuli mainly control IGF-I secretion in the local microenvironment of a particular organ (paracrine activities), while blood IGF-I concentration is the most important variable for balanced systemic growth (endocrine activities).
B. Clinical applications • Growth retardation: Growth retardation may be due to several causes, among which deficient GH production (hypopituitarism), which is associated with low IGF-I blood levels. Because of the difficulties to get interpretable results from GH measurements (by dynamic multiple or stimulation tests), the determination of the stable IGF-I concentration in plasma is often considered as a simple screening test to evaluation \"GH impregnation\" of the patient before deciding more extensive investigations. In several clinical situations with impaired growth, low IGF-I levels may be observed despite normal or high GH production (i.e. malnutrition, chronic diseases states, some genetic dwarfs like Pygmies,..). Interestingly, children with discrete GH neuro-secretery dysfunction may display low IGF-I values despite normal GH levels by conventional testing. The results of IGF-I assay must be interpreted cautiously by considering the normal variations of IGF-I during childhood and adolescence (see Rosenfeld et al). • Acromegaly: IGF-I levels are elevated in acromegaly (excess production of GH) and may serve as an indicator of disease severity. Results are more readily interpreted because the normal values are more easily defined in adults. IGF-I measurements are also useful to monitor treatment. • Research: The IGF-I EASIA kit is an invaluable tool to study the modifications of this growth factor during physiologic (i.e. pregnancy) or pathologic (i.e. diabetes) situations, and the local regulation of IGF-I production in relation to its paracrine and autocrine activities (wound healing, organ regeneration, neoplastic growth, foetal development, gonadal regulation, etc).
Principles of the method The I GF1-EASIA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on breakable microtiterplates.
In the present kit, has introduced a pre-treatment step in order to improve the clinical performance of the assay. It is well established that the binding proteins interfere with the EASIA assay for IGF1.The pre-treatment step used by is the acid-ethanol procedure of Daughaday et al.(8)
A fixed amount of IGF1-labelled with horseradish peroxidase (HRP), compete with unlabelled IGF1 present in the calibrators ,controls and samples for a limited number of binding sites on a specific antibody. After 1 hour incubation at room temperature, the microtiterplate is washed to stop the competition reaction. The Chromogenic solution (TMB ) is added and incubated for 30 min. The reaction is stopped with the addition of Stop Solution and the microtiterplate is then read at the appropriate wavelength. The amount of substrate turnover is determined colourimetrically by measuring the absorbance, which is inversely proportional to the IGF1 concentration.
A calibration curve is plotted and IGF1 concentration in samples is determined by interpolation from the calibration curve.
Storage and Stability • Before opening or reconstitution, all kit components are stable until the expiry date, indicated on the vial label, if kept at 2 to 8°C.
• Unused strips must be stored, at 2°C-8°C, in a sealed bag containing a desiccant until expiration date.
After reconstitution, calibrators and controls are stable for one week at 2 to 8°C. For longer storage periods, aliquots should be made and kept at -20°C for maximum 3 months. Avoid successive freezing and thawing
The concentrated Wash Solution is stable at room temperature until expiration date.
• Freshly prepared Working Wash solution should be used on the same day.
The Working IGF1-HRP conjugate is stable for 4 hours at room temperature, avoid direct sunlight.
Alterations in physical appearance of kit reagents may indicate instability or deterioration.
Product Name Insulin Growth Factor-1.
Citation Zhong D, Xiong L, Liu T, Liu X, Liu X, Chen J, Sun SY, Khuri FR, Zong Y, Zhou Q, Zhou W. The glycolytic inhibitor 2-deoxyglucose activates multiple prosurvival pathways through IGF1R. J Biol Chem. 2009 Aug 28;284(35):23225-33. Epub 2009 Jul 1. PubMed PMID: 19574224; PubMed Central PMCID: PMC2749096.
Experiment Name Active free IGF-1 ELISA.
Experiment Background 1. Diansheng et. al. discovered that 2-DG treatments disrupted the binding between insulin-like growth factor 1 (IGF-1) and IGF-binding protein 3 (IGFBP3) so that the free form of IGF-1 could be released from the IGF-1.IGFBP3 complex to activate IGF-1 receptor (IGF1R) signaling.
2. 2-deoxyglucose (2-DG) is well characterized as a glycolytic inhibitor.
Experiment Steps 1. Incubate 100 pg of free IGF-1 with 20 ng of IGFBP3 in a 10-ul volume at 4°C overnight to form the IGF-1.IGFBP3 complex.
2. Treat this complex to various concentrations of 2-DG for 15 min at room temperature.
3. Determine the active free IGF-1 ELISA kit using a protocol provided by the manufacturer.
Function The insulin-like growth factors, isolated from plasma, are structurally and functionally related to insulin but have a much higher growth-promoting activity.
Subcellular Location Secreted.
Similarity Belongs to the insulin family.

Additional Information

Name Insulin Growth Factor-1 ELISA Kit
Related Product Names Insulin Growth Factor-1 ELISA Kit; IGF-I; Somatomedin Igf-1Insulin Growth Factor-1IGF-1
Molecular Weight 17093
Datasheets / Downloads GWB-6F09CA Datasheet
Swiss Prot Number P05017
Intended Use Research Use Only