Mouse RAGE Quantikine ELISA Kit

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Mouse RAGE Quantikine ELISA Kit Summary

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), Tissue Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (50 uL)
Sensitivity
4.81 pg/mL
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and recombinant mouse RAGE.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Mouse RAGE Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse RAGE levels in cell culture supernates, tissue lysates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse RAGE and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse RAGE accurately. Results obtained using natural mouse RAGE showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse RAGE.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 66.1 236 518 68.8 242 559
Standard Deviation 5.02 14.5 24.3 5.44 13.4 32.6
CV% 7.6 6.1 4.7 7.9 5.5 5.8

Recovery

The recovery of mouse RAGE spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 108 94-120
EDTA Plasma (n=4) 101 87-118
Heparin Plasma (n=4) 111 96-120
Serum (n=4) 107 90-117
Tissue Lysates (n=4) 103 91-120
Urine (n=4) 109 96-120

Linearity

To assess the linearity of the assay, samples containing high concentrations of mouse RAGE were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Mouse RAGE ELISA Linearity

Scientific Data

Mouse RAGE ELISA Standard Curve

Product Datasheets

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Preparation and Storage

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

Background: RAGE/AGER

RAGE (Receptor for Advanced Glycation End product) is a transmembrane glycoprotein that binds advanced glycation end products (AGEs), beta-amyloid peptides, HMGB1/Amphoterin, and several S100 family proteins. AGEs are adducts formed by the non-enzymatic glycation and oxidation of proteins and lipids. A soluble form can also be generated by MMP-mediated shedding. RAGE is expressed in the CNS during development as well as in adult endothelial cells, smooth muscle cells, pericytes, monocytes, and neurons. It is locally upregulated in vascular inflammation (e.g. diabetes, atherosclerosis, vascular injury, Alzheimer’s disease). At these sites, RAGE binding to S100A1, EN-RAGE/S100A12, or S100B induces inflammatory immune cell adhesion and infiltration as well as vascular smooth muscle proliferation, neointimal expansion, atherosclerotic plaque development, and transport of A-beta into the cerebrospinal fluid. In cancer, RAGE binding to HMGB1, S100A8, or S100A9 promotes tumor growth and metastasis in addition to inflammatory cell infiltration.

To view our complete solutions for RAGE research, visit bio-techne.com.

Long Name:
Receptor for Advanced Glycation End Products
Entrez Gene IDs:
177 (Human); 11596 (Mouse); 81722 (Rat); 403168 (Canine)
Alternate Names:
advanced glycosylation end product-specific receptor; AGER; RAGE isoform delta; RAGE isoform sRAGE-delta; RAGE; Receptor for advanced glycosylation end products; receptor for advanced glycosylation end-products; SCARJ1
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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 3 times for a total of 4 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 4 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 for Mouse RAGE Quantikine ELISA Kit

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. Biological Effects of Corticosteroids on Pneumococcal Pneumonia in Mice and Humans
    Authors: Taenaka, H;Wick, KD;Sarma, A;Matsumoto, S;Ghale, R;Fang, X;Maishan, M;Gotts, JE;Langelier, CR;Calfee, CS;Matthay, MA;
    Research square
    Species: Mouse
    Sample Types: BALF
  2. Effects of sevoflurane on lung epithelial permeability in experimental models of acute respiratory distress syndrome
    Authors: Zhai, R;Lenga Ma Bonda, W;Leclaire, C;Saint-Béat, C;Theilliere, C;Belville, C;Coupet, R;Blondonnet, R;Bouvier, D;Blanchon, L;Sapin, V;Jabaudon, M;
    Journal of translational medicine
    Species: Mouse
    Sample Types: BALF, Cell Culture Supernates
  3. NETs Promote Inflammatory Injury by Activating cGAS-STING Pathway in Acute Lung Injury
    Authors: J Zhao, N Zhen, Q Zhou, J Lou, W Cui, G Zhang, B Tian
    International Journal of Molecular Sciences, 2023-03-07;24(6):.
    Species: Mouse
    Sample Types: Serum
  4. Impairing RAGE signaling promotes survival and limits disease pathogenesis following SARS-CoV-2 infection in mice
    Authors: F Jessop, B Schwarz, D Scott, LM Roberts, E Bohrnsen, JR Hoidal, CM Bosio
    JCI Insight, 2022-01-25;7(2):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. Fermented rice bran supplementation attenuates chronic colitis-associated extraintestinal manifestations in female C57BL/6N mice
    Authors: J Islam, AZ Agista, K Watanabe, T Nochi, H Aso, Y Ohsaki, T Koseki, M Komai, H Shirakawa
    The Journal of nutritional biochemistry, 2021-09-10;0(0):108855.
    Species: Mouse
    Sample Types: Serum
  6. ADAM10 attenuates the development of abdominal aortic aneurysms in a mouse model
    Authors: Q Renfeng, C Shuxiao, G Peixian, L Kun, F Xuedong, Y Hai, W Xuejun, L Gang
    Molecular Medicine Reports, 2021-09-07;24(5):.
    Species: Mouse
    Sample Types: Serum
  7. Soluble Receptor for Advanced Glycation End-products regulates age-associated Cardiac Fibrosis
    Authors: F Scavello, F Zeni, G Milano, F Macrì, S Castiglion, E Zuccolo, A Scopece, G Pezone, CC Tedesco, P Nigro, G Degani, E Gambini, F Veglia, L Popolo, G Pompilio, GI Colombo, ME Bianchi, A Raucci
    International journal of biological sciences, 2021-06-11;17(10):2399-2416.
    Species: Mouse
    Sample Types: Serum
  8. Inhibition of the Receptor for Advanced Glycation End Products Enhances the Cytotoxic Effect of Gemcitabine in Murine Pancreatic Tumors
    Authors: P Swami, KA O'Connell, S Thiyagaraj, A Crawford, P Patil, P Radhakrish, S Shin, TC Caffrey, J Grunkemeye, T Neville, SW Vetter, MA Hollingswo, E Leclerc
    Biomolecules, 2021-04-01;11(4):.
    Species: Mouse
    Sample Types: Cell Lysates
  9. Alteration of oxidative-stress and related marker levels in mouse colonic tissues and fecal microbiota structures with chronic ethanol administration: Implications for the pathogenesis of ethanol-related colorectal cancer
    Authors: H Ohira, A Tsuruya, D Oikawa, W Nakagawa, R Mamoto, M Hattori, T Waki, S Takahashi, Y Fujioka, T Nakayama
    PLoS ONE, 2021-02-12;16(2):e0246580.
    Species: Mouse
    Sample Types: Tissue Homogenates
  10. Change of surfactant protein D and A after renal ischemia reperfusion injury
    Authors: IM Imtiazul, R Asma, JH Lee, NJ Cho, S Park, HY Song, HW Gil
    PLoS ONE, 2019-12-26;14(12):e0227097.
    Species: Mouse
    Sample Types: Serum
  11. RAGE inhibition reduces acute lung injury in mice
    Authors: R Blondonnet, J Audard, C Belville, G Clairefond, J Lutz, D Bouvier, L Roszyk, C Gross, M Lavergne, M Fournet, L Blanchon, C Vachias, C Damon-Soub, V Sapin, JM Constantin, M Jabaudon
    Sci Rep, 2017-08-03;7(1):7208.
    Species: Mouse
    Sample Types: Tissue Homogenates
  12. The Mouse-Specific Splice Variant mRAGE_v4 Encodes a Membrane-Bound RAGE That Is Resistant to Shedding and Does Not Contribute to the Production of Soluble RAGE
    PLoS ONE, 2016-09-21;11(9):e0153832.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. Mouse RAGE Variant 4 Is a Dominant Membrane Receptor that Does Not Shed to Generate Soluble RAGE
    PLoS ONE, 2016-09-21;11(9):e0153657.
    Species: Human, Mouse
    Sample Types: Cell Culture Supernates, Serum

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