Mouse IGFBP-3 Quantikine ELISA Kit

  (7 citations)     
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Product Details
Assay Procedure
Citations (7)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), EDTA Plasma (10 uL)
  • Sensitivity
    16 pg/mL
  • Assay Range
    78.1 - 5,000 pg/mL (Cell Culture Supernates, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse IGFBP-3. This kit detects full-length and fragmented mouse IGFBP-3. It also detects IGF-I, IGF-II, IGF-I/ALS, and IGF-II/ALS complexed IGFBP-3.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Mouse IGFBP-3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IGFBP-3 in cell culture supernates and plasma. It contains NS0-expressed recombinant mouse IGFBP-3 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IGFBP-3 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse IGFBP-3.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 280 628 2653 283 602 2648
Standard Deviation 13.5 25.1 148 23.4 44.4 156
CV% 4.8 4 5.6 8.3 7.4 5.9

Recovery

The recovery of mouse IGFBP-3 spiked to three levels throughout the range of the assay in the cell culture supernate samples was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 100 90-109
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse IGFBP-3 in each matrix were diluted with Calibrator Diluent and assayed.
 IGFBP-3 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IGFBP-3
The superfamily of insulin-like growth factor (IGF) binding proteins include the six high-affinity IGF binding proteins (IGFBP) and at least four additional low-affinity binding proteins referred to as IGFBP related proteins (IGFBP-rP). All IGFBP superfamily members are cysteine-rich proteins with conserved cysteine residues, which are clustered in the amino- and carboxy-terminal thirds of the molecule. IGFBPs modulate the biological activities of IGF proteins. Some IGFBPs may also have intrinsic bioactivity that is independent of their ability to bind IGF proteins. Post-translational modifications of IGFBP, including glycosylation, phosphorylation and proteolysis, have been shown to modify the affinities of the binding proteins to IGF. ALS (Acid Labile Subunit) is a liver-derived protein that exists in a ternary complex with Insulin-like Growth Factor (IGF)-binding Protein-3 (IGFBP-3) or IGFBP-5, and either IGF-I or IGF-II. ALS increases the half-life of IGF/IGFBP complexes in circulation.
  • Long Name:
    Insulin-like Growth Factor Binding Protein 3
  • Entrez Gene IDs:
    3486 (Human); 16009 (Mouse)
  • Alternate Names:
    acid stable subunit of the 140 K IGF complex; binding protein 29; binding protein 53; growth hormone-dependent binding protein; IBP-3; IBP3BP-53; IGF-binding protein 3; IGFBP3; IGFBP-3; insulin-like growth factor binding protein 3; insulin-like growth factor-binding protein 3
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

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.

7 Citations: Showing 1 - 7
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Species
Sample Type
  1. Effects of Intestinal Microbiota on Brain Development in Humanized Gnotobiotic Mice
    Authors: J Lu, L Lu, Y Yu, J Cluette-Br, CR Martin, EC Claud
    Sci Rep, 2018;8(1):5443.
    Species: Mouse
    Sample Type: Serum
  2. EGFR controls bone development by negatively regulating mTOR-signaling during osteoblast differentiation
    Authors: M Linder, M Hecking, E Glitzner, K Zwerina, M Holcmann, L Bakiri, MG Ruocco, J Tuckermann, G Schett, EF Wagner, M Sibilia
    Cell Death Differ., 2018;0(0):.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  3. Glucose intolerance in aging male IGFBP-3 transgenic mice: differential effects of human IGFBP-3 and its mutant IGFBP-3 devoid of IGF binding ability.
    Authors: Nguyen K, Yao X, Erickson A, Mishra S, Nyomba B
    Endocrinology, 2015;156(2):462-74.
    Species: Mouse
    Sample Type: Serum
  4. PAI-1-regulated extracellular proteolysis governs senescence and survival in Klotho mice.
    Authors: Eren M, Boe A, Murphy S, Place A, Nagpal V, Morales-Nebreda L, Urich D, Quaggin S, Budinger G, Mutlu G, Miyata T, Vaughan D
    Proc Natl Acad Sci U S A, 2014;111(19):7090-5.
    Species: Mouse
    Sample Type: Plasma
  5. Fish oil slows prostate cancer xenograft growth relative to other dietary fats and is associated with decreased mitochondrial and insulin pathway gene expression.
    Authors: Lloyd J, Masko E, Wu C, Keenan M, Pilla D, Aronson W, Chi J, Freedland S
    Prostate Cancer Prostatic Dis, 2013;16(4):285-91.
    Species: Mouse
    Sample Type: Serum
  6. Deletion of beta-adrenergic receptor 1, 2, or both leads to different bone phenotypes and response to mechanical stimulation.
    Authors: Pierroz DD, Bonnet N, Bianchi EN, Bouxsein ML, Baldock PA, Rizzoli R, Ferrari SL
    J. Bone Miner. Res., 2012;27(6):1252-62.
    Species: Mouse
    Sample Type: Serum
  7. Sexual dimorphism in cortical bone size and strength but not density is determined by independent and time-specific actions of sex steroids and IGF-1: evidence from pubertal mouse models.
    Authors: Callewaert F, Venken K, Kopchick JJ, Torcasio A, van Lenthe GH, Boonen S, Vanderschueren D
    J. Bone Miner. Res., 2010;25(3):617-26.
    Species: Mouse
    Sample Type: Serum

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