Overview

  • Product name

    Rat RAGE ELISA Kit
    See all RAGE kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 2.6%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 2%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    6.9 pg/ml
  • Range

    31.25 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 90 82.4% - 98.8%
    Serum 106.6 100.5% - 116.7%
    Cell culture media 108.3 99.1% - 117.3%
    Citrate Plasma 103.2 95.3% - 112.8%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    Abcam’s RAGE in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of RAGE protein in rat serum, plasma, urine, cell culture supernatant, 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 – 6.9 pg/mL
    Samples diluted in 1X Cell Extraction Buffer PTR – 13.1 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

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Rat RAGE Capture Antibody 1 x 600µl
    10X Rat RAGE Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Plate Seals 1 unit
    Rat RAGE Lyophilized Recombinant Protein 2 vials
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Mediates interactions of advanced glycosylation end products (AGE). These are nonenzymatically glycosylated proteins which accumulate in vascular tissue in aging and at an accelerated rate in diabetes. 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. Interaction with S100A12 on endothelium, mononuclear phagocytes, and lymphocytes triggers cellular activation, with generation of key proinflammatory mediators. Interaction with S100B after myocardial infarction may play a role in myocyte apoptosis by activating ERK1/2 and p53/TP53 signaling (By similarity). Receptor for amyloid beta peptide. 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 intraneuronal space.
  • Tissue specificity

    Endothelial cells.
  • Sequence similarities

    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt
  • Alternative names

    • Advanced glycosylation end product-specific receptor
    • Ager
    • DAMA 358M23.4
    • MGC2235
    • MGC22357
    • RAGE_HUMAN
    • Receptor for advanced glycation end products
    • Receptor for advanced glycosylation end products
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab202409 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.

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

  • The concentrations of RAGE were measured in duplicates, interpolated from the RAGE standard curve and corrected for sample dilution. Note that 1X Diluted serum samples were 10X pre-diluted, 1X Diluted plasma (citrate) samples were 5X pre-diluted, 1X Diluted urine samples were neat. The interpolated, dilution factor-corrected values are plotted in ng of RAGE per mL of neat biological fluid (mean +/- SD, n=2).

Protocols

References

This product has been referenced in:

  • An K  et al. Spinal PKC activation - Induced neuronal HMGB1 translocation contributes to hyperalgesia in a bone cancer pain model in rats. Exp Neurol 303:80-94 (2018). Read more (PubMed: 29428215) »
See 1 Publication for this product

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