Overview

  • Product name

    Mouse RAGE ELISA Kit
    See all RAGE kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 3.3%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 5.9%
  • Sample type

    Cell culture supernatant, Urine, Serum, Cell culture extracts, Tissue Extracts, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    6.6 pg/ml
  • Range

    62.5 pg/ml - 4000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 90.4 88.3% - 93.3%
    Serum 91.7 83.1% - 97.9%
    Cell culture media 91.8 89.8% - 95.4%
    Citrate Plasma 100.8 93.6% - 116.9%
    Goat Serum 98.7 88.9% - 105.2%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rabbit, Goat, Guinea pig, Cow, Dog, Human, Pig
  • Product overview

    RAGE in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of RAGE protein in mouse serum, plasma, urine, cell culture supernatant and cell and tissue extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:


    Samples diluted in Sample Diluent NS – 15.2 pg/mL
    Samples diluted in 1X Cell Extraction Buffer PTR – 6.6 pg/mL

  • Notes

    RAGE mediates interactions of advanced glycosylation end products (AGE). These are non-enzymatically glycosylated proteins which accumulate in vascular tissue in aging and at an accelerated rate in diabetes. RAGE acts as a mediator of both acute and chronic vascular inflammation in conditions such as atherosclerosis and in particular as a complication of diabetes. AGE/RAGE signaling plays an important role in regulating the production/expression of TNF-alpha, oxidative stress, and endothelial dysfunction in type 2 diabetes. RAGE interaction with S100A12 on endothelium, mononuclear phagocytes, and lymphocytes triggers cellular activation, with generation of key pro-inflammatory mediators. RAGE may be a receptor for amyloid beta peptide. RAGE contributes to the translocation of amyloid-beta peptide (ABPP) across the cell membrane from the extracellular to the intracellular space in cortical neurons. ABPP-initiated RAGE signaling, especially stimulation of p38 mitogen-activated protein kinase (MAPK), has the capacity to drive a transport system delivering ABPP as a complex with RAGE to the intra-neuronal space. RAGE-dependent signaling in microglia contributes to neuroinflammation, amyloid accumulation, and impaired learning/memory in a mouse model of Alzheimer disease.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab197745 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Tissue cells were cultured for 5 days and after appropriate dilution the cell culture supernatant samples were analyzed with this kit. Interpolated concentrations of RAGE adjusted for sample dilution are graphed in ng of RAGE per mL of supernatant (mean +/- SD, n = 2).

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Tissue extracts at two different dilutions were analyzed with this kit. Interpolated concentrations of RAGE adjusted for sample dilution are graphed in ng of RAGE per mg of extract (mean +/- SD, n = 2).

Protocols

References

ab197745 has not yet been referenced specifically in any publications.

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