Human RAGE DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1145
Ancillary Products Available
Human RAGE ELISA Standard Curve
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Product Details
Procedure
Citations (9)
FAQs
Supplemental Products
Reviews (5)

Human RAGE 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 human RAGE. 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 ELISA.

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


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 equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

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), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

 

Data Example

Human RAGE 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: 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) 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 each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

    Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
     
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

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 (as recommended in the product datasheet), 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 Human RAGE 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.

9 Citations: Showing 1 - 9
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  1. Modulation of soluble receptor for advanced glycation end-products (RAGE) isoforms and their ligands in healthy aging
    Authors: F Scavello, F Zeni, CC Tedesco, E Mensà, F Veglia, AD Procopio, AR Bonfigli, F Olivieri, A Raucci
    Aging (Albany NY), 2019;11(6):1648-1663.
    Species: Human
    Sample Types: Serum
  2. Regulation of RAGE ectodomain shedding and its role in cell function
    Authors: A Braley, T Kwak, J Jules, E Harja, R Landgraf, BI Hudson
    J Biol Chem, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Expression of RAGE and HMGB1 in thymic epithelial tumors, thymic hyperplasia and regular thymic morphology.
    Authors: Moser B, Janik S, Schiefer A, Mullauer L, Bekos C, Scharrer A, Mildner M, Renyi-Vamos F, Klepetko W, Ankersmit H
    PLoS ONE, 2014;9(4):e94118.
    Species: Human
    Sample Types: Serum
  4. Circulating endothelial progenitor cells and large artery structure and function in young subjects with uncomplicated type 1 diabetes.
    Authors: Palombo C, Kozakova M, Morizzo C, Gnesi L, Barsotti M, Spontoni P, Massart F, Salvi P, Balbarini A, Saggese G, Di Stefano R, Federico G
    Cardiovasc Diabetol, 2011;10(0):88.
    Species: Human
    Sample Types: Plasma
  5. Circulating soluble receptor for advanced glycation end-product levels are decreased in patients with calcific aortic valve stenosis.
    Authors: Basta G, Corciu AI, Vianello A, Del Turco S, Foffa I, Navarra T, Chiappino D, Berti S, Mazzone A
    Atherosclerosis, 2010;210(2):614-8.
    Species: Human
    Sample Types: Plasma
  6. High plasma levels of the soluble receptor for advanced glycation endproducts in patients with symptomatic carotid atherosclerosis.
    Authors: Basta G, Castagnini M, Del Turco S, Epistolato MC, Righini P, Sangiorgi GM, De Caterina R, Tanganelli P
    Eur. J. Clin. Invest., 2009;39(12):1065-72.
    Species: Human
    Sample Types: Plasma
  7. Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases.
    Authors: Hsu HY, Wittemann S, Schneider EM, Weiss M, Joos TO
    Med Eng Phys, 2008;30(8):976-83.
    Species: Human
    Sample Types: Plasma
  8. Distribution of the receptor for advanced glycation end products in the human male reproductive tract: prevalence in men with diabetes mellitus.
    Authors: Mallidis C, Agbaje I, Rogers D, Glenn J, McCullough S, Atkinson AB, Steger K, Stitt A, McClure N
    Hum. Reprod., 2007;22(8):2169-77.
    Species: Human
    Sample Types: Seminal Plasma
  9. Lung epithelial injury markers are not influenced by use of lower tidal volumes during elective surgery in patients without preexisting lung injury.
    Authors: Determann RM, Wolthuis EK, Choi G, Bresser P, Bernard A, Lutter R, Schultz MJ
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007;294(2):L344-50.
    Species: Human
    Sample Types: BALF

FAQs

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Reviews for Human RAGE DuoSet ELISA

Average Rating: 4.6 (Based on 5 Reviews)

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Human RAGE DuoSet ELISA
By Anonymous on 10/11/2018
Application: Sample Tested: Serum and Plasma

Human RAGE DuoSet ELISA
By Anonymous on 10/08/2018
Application: Sample Tested: EDTA Plasma

Human RAGE DuoSet ELISA
By Anonymous on 12/20/2017
Application: Sample Tested: EDTA Plasma

Human RAGE DuoSet ELISA
By Anonymous on 07/11/2017
Application: Sample Tested: EDTA Plasma

We run human EDTA-plasma samples with this kit at an 1:5 dilution. The observed inter coefficient of variation was a bit higher than preferred.


Human RAGE DuoSet ELISA
By Anonymous on 05/22/2017
Application: Sample Tested: Lung tissue