ELISA Kit for Insulin Like Growth Factor Binding Protein 3 (IGFBP3)

BP53; IBP3; Growth Hormone-Dependent Binding Protein; Acid Stable Subunit Of The 140 K IGF Complex

Specificity

This assay has high sensitivity and excellent specificity for detection of Insulin Like Growth Factor Binding Protein 3 (IGFBP3).
No significant cross-reactivity or interference between Insulin Like Growth Factor Binding Protein 3 (IGFBP3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Insulin Like Growth Factor Binding Protein 3 (IGFBP3) and the recovery rates were calculated by comparing the measured value to the expected amount of Insulin Like Growth Factor Binding Protein 3 (IGFBP3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-104 98
EDTA plasma(n=5) 90-99 94
heparin plasma(n=5) 97-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Insulin Like Growth Factor Binding Protein 3 (IGFBP3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Insulin Like Growth Factor Binding Protein 3 (IGFBP3) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Insulin Like Growth Factor Binding Protein 3 (IGFBP3) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 89-97% 91-105% 89-101% 78-96%
EDTA plasma(n=5) 79-96% 82-94% 88-104% 83-90%
heparin plasma(n=5) 91-99% 85-98% 80-97% 99-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Molecular Biology Reports Characterization of the specific and sustained GH1 expression induced by rAAV2/1 in normal adult male rats PubMed: 20204528
Endocrinology Role of IGFBP-3 in the Regulation of β-Cell Mass during Obesity: Adipose Tissue/β-Cell Cross Talk Endo: source
Nutrition and Cancer A Pilot Study Comparing the Effect of Flaxseed, Aromatase Inhibitor, and the Combination on Breast Tumor Biomarkers Tandfonline: Source
Dig Dis Sci The Protective Effects of Exclusive Enteral Nutrition Formulas on Growth Factor Expression and the Proximal Tibial Epiphyseal Growth Plate in a TNBS-Induced IBD Rat Model PubMed: 25708898
Animal. Effects of a dietary crude fibre concentrate on growth in weaned piglets. pubmed:28318464
PLoS One Altered insulin-like growth factor-2 signaling is associated with psychopathology and cognitive deficits in patients with schizophrenia Pubmed: 32191705
Int J Environ Res Public Health Association of Salivary IGF and IGF/IGFBP-3 Molar Ratio with Cervical Vertebral Maturation Stages from Pre-Adolescent to Post-Adolescent Transition Period—A … Pubmed:35564566
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