Description

Nori Monkey RAGE ELISA Kit Summary

Alternative names for RAGE: Receptor for advanced glycation endproducts), AGER

Alternative names for monkey: various species of monkey

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

RAGE (receptor for advanced glycation endproducts), also called AGER, is a 35 kilodalton transmembrane receptor of the immunoglobulin super family which was first characterized in 1992 by Neeper et al.^[1]  The interaction between RAGE and its ligands is thought to result in pro-inflammatory gene activation.^[2] [3] Due to an enhanced level of RAGE ligands in diabetes or other chronic disorders, this receptor may have a causative effect in a range of inflammatory diseases such as diabetic complications, Alzheimer’s disease and even some tumors. Isoforms of the RAGE protein, which lack the transmembrane and the signaling domain (commonly referred to as soluble RAGE or sRAGE) may counteract the detrimental action of the full-length receptor. RAGE has been linked to several chronic diseases, which are thought to result from vascular damage. RAGE itself is upregulated by NF-κB. Given a condition in which there is a large amount of RAGE ligands (e.g. AGE in diabetes or amyloid-β-protein in Alzheimer’s disease) this establishes a positive feed-back cycle, which leads to chronic inflammation. This chronic condition is then believed to alter the micro- and macrovasculature, resulting in organ damage or even organ failure.^[4] RAGE is expressed at the highest levels in the lung compared to other tissues, in particular in alveolar epithelial type I cells, and is lost in Idiopathic pulmonary fibrosis indicating that expression and regulation of RAGE in the pulmonary system differs from that in the vascular system. Blockade/knockdown of RAGE resulted in impaired cell adhesion, and increased cell proliferation and migration^[5]

References

1. Neeper M, et al. (1992). The Journal of Biological Chemistry. 267 (21): 14998–5004.
2. Bierhaus A, et al. (2001). Diabetes. 50 (12): 2792–808.
3. Gasparotto, J; et al. (2017). J Biol Chem. 293: 226–244.
4. Gasparotto, J; et al. (2019). Mol Neurobiol. 56(5): 3079–3089.
5. Queisser MA, (2008). American J Respiratory Cell and Molecular Biology. 39 (3): 337–45.

Product Citations