Mouse/Rat IGF-I/IGF-1 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY791
Ancillary Products Available
Mouse / Rat IGF-I ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (13)
FAQs
Supplemental Products
Reviews (2)

Mouse/Rat IGF-I/IGF-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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 mouse 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.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY004), or 5% Tween 20 in PBS, 7.2-7.4, 0.2 μm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

Data Example

Mouse / Rat IGF-I ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IGF-I/IGF-1

Insulin-like Growth Factor 1 (IGF-1), also known as somatomedin C, is a member of the insulin superfamily. It was originally discovered as a mediator of growth hormone actions on somatic cell growth, but has also been shown to be an important regulator of cell metabolism, differentiation and survival. IGF-1 is synthesized as a preproprotein that is proteolytically cleaved to generate the mature protein linked by three disulfide bonds. Mature IGF-1 is highly conserved among large mammals, with 100% sequence identity between the human, bovine, porcine, equine, and canine proteins. 


Mature mouse IGF-1 is a non-glycosylated, 70 amino acid (aa) secreted polypeptide that is derived from either a 153 aa or a 159 aa preproprotein. It shares 99% and 94% aa sequence identity with rat and human IGF-1, respectively. IGF-1 is synthesized in the liver and other tissues. It is found in blood and other body fluids as a complex with specific high affinity IGF binding proteins (IGFBP-1 to -6). The IGFBPs are expressed in specific patterns during development. They are modulators of IGF actions, which control IGF bioavailability to specific cell-surface receptors. Their functions are further regulated by IGFBP proteases, which proteolytically cleave the IGFBPs to lower the affinity with which they bind IGFs and increase IGF bioavailability. Some IGFBPs also have IGF-Independent effects on cell functions. IGF-Independent circulates primarily as a ternary complex with IGFBP-3 or IGFBP-5 and the acid-labile subunit (ALS). Some IGF-1 is also present in binary complexes with other IGFBPs. Whereas the ternary complexes are generally restricted to the vasculature, the binary complexes freely enter the tissues. 

IGF-1 actions are mediated by two ubiquitously expressed receptor tyrosine kinases: IGF-I R and Insulin R/CD220. IGF-I R and Insulin R are disulfide-linked heterotetrameric complexes that consist of two alpha and two beta subunits. For both of these receptors, the prepro proteins are cleaved to produce extracellular alpha subunits which contain a cysteine-rich region and ligand-binding fibronectin type III (FN-III) domains, and beta subunits which contain an extracellular FN-III domain, transmembrane, and cytoplasmic tyrosine kinase domains. A hybrid complex containing one IGF-I R and one Insulin R also serves as a functional high affinity receptor for IGF-1. IGF-I R-Insulin R hybrids respond primarily to IGF-1, potentially downregulating the cellular response to Insulin. IGF signaling is also modulated by IGF binding proteins and the scavenger receptor, IGF-I I R.

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

Assay Procedure

GENERAL ELISA PROTOCOL


Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL of Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

Citations for Mouse/Rat IGF-I/IGF-1 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. Proteasome Activation by Insulin-like Growth Factor-1/Nuclear Factor Erythroid 2-related Factor 2 Signaling Promotes Exercise-induced Neurogenesis
    Authors: X Niu, Y Zhao, N Yang, X Zhao, W Zhang, X Bai, A Li, W Yang, L Lu
    Stem Cells, 2019;0(0):.
    Species: Mouse
    Sample Types: Serum
  2. Effects of IGF-1 isoforms on muscle growth and sarcopenia
    Authors: F Ascenzi, L Barberi, G Dobrowolny, A Villa Nova, C Nicoletti, E Rizzuto, N Rosenthal, BM Scicchitan, A Musarò
    Aging Cell, 2019;0(0):e12954.
    Species: Mouse
    Sample Types: Serum
  3. Inflammation in the hippocampus affects IGF1 receptor signaling and contributes to neurological sequelae in rheumatoid arthritis
    Authors: KME Andersson, C Wasén, L Juzokaite, L Leifsdotti, MC Erlandsson, ST Silfverswä, A Stokowska, M Pekna, M Pekny, K Olmarker, RA Heckemann, M Kalm, MI Bokarewa
    Proc. Natl. Acad. Sci. U.S.A., 2018;0(0):.
    Species: Mouse
    Sample Types: Serum
  4. Epigenetic Restoration of Fetal-like IGF1 Signaling Inhibits Leukemia Stem Cell Activity
    Authors: V Giambra, S Gusscott, D Gracias, R Song, SH Lam, P Panelli, K Tyshchenko, CE Jenkins, C Hoofd, A Lorzadeh, A Carles, M Hirst, CJ Eaves, AP Weng
    Cell Stem Cell, 2018;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Mycobacterium leprae-induced Insulin-like Growth Factor I attenuates antimicrobial mechanisms, promoting bacterial survival in macrophages
    Authors: L R Batista-Si
    Sci Rep, 2016;6(0):27632.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. Hypothalamic gene transfer of BDNF inhibits breast cancer progression and metastasis in middle age obese mice.
    Authors: Liu, Xianglan, McMurphy, Travis, Xiao, Run, Slater, Andrew, Huang, Wei, Cao, Lei
    Mol Ther, 2014;22(7):1275-84.
    Species: Mouse
    Sample Types: Serum
  7. The lung inflammation and skeletal muscle wasting induced by subchronic cigarette smoke exposure are not altered by a high-fat diet in mice.
    Authors: Hansen, Michelle, Chen, Hui, Jones, Jessica, Langenbach, Shenna Y, Vlahos, Ross, Gualano, Rosa C, Morris, Margaret, Anderson, Gary P
    PLoS ONE, 2013;8(11):e80471.
    Species: Mouse
    Sample Types: Tissue Homogenates
  8. Interleukin-1 participates in the classical and alternative activation of microglia/macrophages after spinal cord injury.
    Authors: Sato A, Ohtaki H, Tsumuraya T, Song D, Ohara K, Asano M, Iwakura Y, Atsumi T, Shioda S
    J Neuroinflammation, 2012;9(0):65.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. The somatotrope as a metabolic sensor: deletion of leptin receptors causes obesity.
    Authors: Childs GV, Akhter N, Haney A
    Endocrinology, 2011;152(1):69-81.
    Species: Mouse
    Sample Types: Serum
  10. Plasma proteome profiles associated with inflammation, angiogenesis, and cancer.
    Authors: Kelly-Spratt KS, Pitteri SJ, Gurley KE, Liggitt D, Chin A, Kennedy J, Wong CH, Zhang Q, Buson TB, Wang H, Hanash SM, Kemp CJ
    PLoS ONE, 2011;6(5):e19721.
    Species: Mouse
    Sample Types: Plasma
  11. GRP94 is essential for mesoderm induction and muscle development because it regulates insulin-like growth factor secretion.
    Authors: Wanderling S, Simen BB, Ostrovsky O, Ahmed NT, Vogen SM, Gidalevitz T, Argon Y
    Mol. Biol. Cell, 2007;18(10):3764-75.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Mutant SOD1(G93A) microglia are more neurotoxic relative to wild-type microglia.
    Authors: Xiao Q, Zhao W, Beers DR, Yen AA, Xie W, Henkel JS, Appel SH
    J. Neurochem., 2007;102(6):2008-19.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. Protective effects of an anti-inflammatory cytokine, interleukin-4, on motoneuron toxicity induced by activated microglia.
    Authors: Zhao W, Xie W, Xiao Q, Beers DR, Appel SH
    J. Neurochem., 2006;99(4):1176-87.
    Species: Rat
    Sample Types: Cell Culture Supernates

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Reviews for Mouse/Rat IGF-I/IGF-1 DuoSet ELISA

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Mouse/Rat IGF-I DuoSet ELISA
By Anonymous on 11/06/2018
Application: Sample Tested: Serum

Mouse/Rat IGF-I DuoSet ELISA
By Anonymous on 03/31/2018
Application: Sample Tested: Serum