Human IGFBP3 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Insulin-like growth factor (IGF) -binding protein-3 (IGFBP-3) is a major determinant of circulating levels of the IGFs and is clinically useful for the evaluation of GH deficiency and for predicting the response to GH treatment. The circulating level of IGFBP-3 is inversely related to the risk of several common cancers, and that antiproliferative agents such as antiestrogens and retinoids act in part by up-regulating IGFBP-3 gene (IGFBP3) expression. Insulin-like growth factor-binding protein (IGFBP) -3, well characterized as the carrier of insulin-like growth factor (IGF), has been reported to have intrinsic bioactivity that is independent of IGF binding. IGFBP-3 has an IGF-independent, antiproliferative effect in undifferentiated and early differentiated but not in terminally differentiated chondrocytes. IGFBP-3 possesses both growth-inhibitory and potentiating effects on cells that are independent of IGF action and are mediated through specific. IGFBP-3 binding proteins/receptors locate at the cell membrane, cytosol, or nuclear compartments and in the extrIGFBP-3llular matrix. IGFBP3 is also located on chromosome 7. The standard product used in this kit is recombinant human IGFBP-3, consisting of 265 amino acids with the molecular mass of 29kDa. As a result of glycosylation, the molecular mass is 41KDa.

Acid stable subunit of the 140 K IGF complex; Binding protein 29; Binding protein 53; BP 53; BP53; Growth hormone dependent binding protein; IBP 3; IBP3; IGF binding protein 3; IGFBP 3; Insulin Like Growth Factor Binding Protein 3

Cell Biology, Cell Death, Diabetes-associated, Growth Factors/Hormones, Metabolism, Obesity, Protein Phosphorylation, Receptors, Signal Transduction, Tyrosine Phosphatases

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 271 1607 5480 253 1639 5548 Standard deviation 16.53 69.1 350.72 19.73 95.06 438.29 CV (%) 6.1% 4.3% 6.4% 7.8% 5.8% 7.9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 271 1607 5480 253 1639 5548 Standard deviation 16.53 69.1 350.72 19.73 95.06 438.29 CV (%) 6.1% 4.3% 6.4% 7.8% 5.8% 7.9% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 271 1607 5480 253 1639 5548 Mean (pg/ml) 271 1607 5480 253 1639 5548 Standard deviation 16.53 69.1 350.72 19.73 95.06 438.29 Standard deviation 16.53 69.1 350.72 19.73 95.06 438.29 CV (%) 6.1% 4.3% 6.4% 7.8% 5.8% 7.9% CV (%) 6.1% 4.3% 6.4% 7.8% 5.8% 7.9%

IGF-binding proteins prolong the half-life of the IGFs and have been shown to either inhibit or stimulate the growth promoting effects of the IGFs on cell culture. They alter the interaction of IGFs with their cell surface receptors. Also exhibits IGF-independent antiproliferative and apoptotic effects mediated by its receptor TMEM219/IGFBP-3R.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0386-CA Rabbit anti- human IGFBP3 polyclonal antibody (Capture Antibody) 500 μl, 0.3 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0386-DA Biotinylated goat anti- human IGFBP3 polyclonal antibody (Detection Antibody) 500 μl, 2 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0386-ST Lyophilized recombinant human IGFBP3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0386-CA Rabbit anti- human IGFBP3 polyclonal antibody (Capture Antibody) 500 μl, 0.3 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0386-DA Biotinylated goat anti- human IGFBP3 polyclonal antibody (Detection Antibody) 500 μl, 2 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0386-ST Lyophilized recombinant human IGFBP3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0386-CA Rabbit anti- human IGFBP3 polyclonal antibody (Capture Antibody) 500 μl, 0.3 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0386-CA Rabbit anti- human IGFBP3 polyclonal antibody (Capture Antibody) 500 μl, 0.3 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0386-DA Biotinylated goat anti- human IGFBP3 polyclonal antibody (Detection Antibody) 500 μl, 2 μg/mL (recommended dilution 1:100) EZ0386-DA Biotinylated goat anti- human IGFBP3 polyclonal antibody (Detection Antibody) 500 μl, 2 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0386-ST Lyophilized recombinant human IGFBP3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0386-ST Lyophilized recombinant human IGFBP3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.034 0.092 0.16 0.23 0.397 0.691 1.251 1.71 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.034 0.092 0.16 0.23 0.397 0.691 1.251 1.71 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.034 0.092 0.16 0.23 0.397 0.691 1.251 1.71 O.D. 0.034 0.092 0.16 0.23 0.397 0.691 1.251 1.71

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)