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hIGF-I/IGF-1 DuoSet (1 KIT)
品牌： R&D Systems
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Scientific Data
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL of Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Human IGF-I/IGF-1 DuoSet ELISA Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
93.8 - 6,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human IGF-I. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
背景
别名
IBP1,IGF1,IGF-1,IGF1A,IGFI,IGF-I,IGF-IA,IGF-IB,insulin-like growth factor 1 (somatomedin C),insulin-like growth factor 1,insulin-like growth factor I,insulin-like growth factor IA,insulin-like growth factor IB,Mechano growth factor,MGF,Somatomedin A,Somatomedin C,somatomedin-C,Insulin-like Growth Factor I/Insulin-like Growth Factor 1
背景
Background: IGF-I/IGF-1
Insulin-like growth factor 1 (IGF-1, also known as somatomedin C) is a 7.6 kDa, 70 amino acid (aa) polypeptide with three internal disulfide bonds. The sequence of human IGF-1 is identical to that of bovine and porcine IGF-1, and is 70% identical to human IGF2. IGF-1 is a single-chain molecule with about 50% identity to the sequences of the A- and B-chains of human insulin.
IGF-1 is a growth hormone that plays a role in a variety of biological events. It is produced primarily by hepatocytes, serving an endocrine function. It is also produced by many other cells, where it may act in an autocrine or paracrine manner. It binds two tyrosine kinase receptors, the IGF-1 receptor (IGF1R) and the insulin receptor, to initiate downstream events like the AKT and PI3K signal transduction pathways. This triggers cell proliferation and protects cells from apoptosis. IGF-1 also interacts with seven IGF-binding proteins (IGFBP-1 through IGFBP7), influencing IGF-1 binding to IGF1R and increasing IGF-1 half-life to closely regulate IGF-1 signaling. For example, IGFBP-3 binds over 90% of the total IGF in serum in a complex of IGF, IGFBP, and an acid-labile subunit. This ternary complex greatly stabilizes IGF in the circulation, changing the half-life from minutes to hours. Proteases also facilitate IGF-1R binding by cleaving IGFBPs to modify their affinity for IGF or completely eliminate the IGFBP. The interactions of IGF, IGFBP, IGFBP proteases, and IGF receptors are referred to as the IGF axis.
The IGF axis affects many primary physiological and pathological processes, including development, growth, metabolic regulation, tumorigenesis, atherosclerosis, and angiogenesis. Its ability to inhibit apoptosis and stimulate cell growth and proliferation plays a significant role in prenatal development, growth to adulthood, and metabolic control. Serum levels of IGF-1 have been reported to increase from birth to puberty, followed by a slow decline through adulthood. IGF-1 also induces amino acid uptake, protein synthesis, and glucose utilization. In the brain, IGF-1 acts as a neurotrophic factor to promote neurogenesis and neuronal survival. Exercise increases levels of IGF-1 in blood serum, indicating it plays a role as a key mediator of exercise-induced neurogenesis.
Long Name:
Insulin-like Growth Factor I/Insulin-like Growth Factor 1
Entrez Gene IDs:
3479 (Human); 16000 (Mouse); 24482 (Rat)
Alternate Names:
IBP1; IGF1; IGF-1; IGF1A; IGFI; IGF-I; IGF-IA; IGF-IB; insulin-like growth factor 1 (somatomedin C); insulin-like growth factor 1; insulin-like growth factor I; insulin-like growth factor IA; insulin-like growth factor IB; Mechano growth factor; MGF; Somatomedin A; Somatomedin C; somatomedin-C
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