Mouse RAGE Quantikine ELISA Kit

R&D Systems | Catalog # MRG00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Tissue Lysates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Urine (50 µL)

Sensitivity

4.81 pg/mL

Assay Range

31.3-2000 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
View all RAGE/AGER ELISA Kits »

Product Summary for Mouse RAGE Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Mouse

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.

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, EDTA Plasma, Heparin Plasma, Serum, Tissue Lysates, 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 for Mouse RAGE Quantikine ELISA Kit

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 Images for Mouse RAGE Quantikine ELISA Kit

Mouse RAGE ELISA Standard Curve

Mouse RAGE ELISA Standard Curve

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.

Long Name

Receptor for Advanced Glycation End Products

Alternate Names

AGER, SCARJ1

Entrez Gene IDs

177 (Human); 11596 (Mouse); 81722 (Rat); 403168 (Canine)

Gene Symbol

AGER

Additional RAGE/AGER Products

Product Documents for Mouse RAGE Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse RAGE Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Mouse RAGE Quantikine ELISA Kit

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Protocols

View specific protocols for Mouse RAGE Quantikine ELISA Kit (MRG00):

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 100 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   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.

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