Nori Ferret Resistin ELISA Kit Summary

Alternative names for resistin: Adipose tissue-specific secretory factor, ADSF, C/EBP-epsilon-regulated myeloid-specific secreted cysteine-rich protein, XCP1, RETN

Alternative names for ferret: Black-footed ferret, European domestic ferret,

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:

Resistin also known as adipose tissue-specific secretory factor (ADSF) or C/EBP-epsilon-regulated myeloid-specific secreted cysteine-rich protein (XCP1), is a cysteine-rich adipose-derived peptide hormone that is encoded by the RETN gene.^[1] In primates, pigs, and dogs, resistin is secreted by immune and epithelial cells, while, in rodents, it is secreted by adipose tissue. The length of the resistin pre-peptide in human is 108 amino acid residues and in the mouse and rat it is 114 aa. Resistin is an adipose-derived hormone (similar to a cytokine) whose physiologic role has been the subject of much controversy regarding its involvement with obesity and type II diabetes mellitus (T2DM).^[2] Resistin has been shown to cause “high levels of ‘bad’ cholesterol (LDL), increasing the risk of heart disease.  Resistin increases the production of LDL in human liver cells and also degrades LDL receptors in the liver. As a result, the liver is less able to clear ‘bad’ cholesterol from the body. Resistin accelerates the accumulation of LDL in arteries, increasing the risk of heart disease. Resistin adversely impacts the effects of statins, the main cholesterol-reducing drug used in the treatment and prevention of cardiovascular disease.” Resistin has been shown to increase transcriptional events, leading to an increased expression of several pro-inflammatory cytokines including (but not limited to) interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-12 (IL-12), and tumor necrosis factor-α (TNF-α) in an NF-κB-mediated.^[3][4] It has also been demonstrated that resistin upregulates intercellular adhesion molecule-1 (ICAM1), vascular cell-adhesion molecule-1 (VCAM1) and chemokine (C-C motif) ligand 2 (CCL2), all of which are occupied in chemotactic pathways involved in leukocyte recruitment to sites of infection.^[5] Resistin itself can be upregulated by interleukins and also by microbial antigens such as lipopolysaccharide,^[6] which are recognized by leukocytes. Resistin may be a link between inflammation and insulin resistance.^[7]

References

1. Wang H, et al. (2002). J. Clin. Endocrinol. Metab. 87 (6): 2520–4.
2. Lazar MA (2007). Horm. Metab. Res. 39 (10): 710–6.
3. Milan Get al. (2002). Obes. Res. 10 (11): 1095–103.
4. Silswal N, et al. (2005). Biochem. Biophys. Res. Commun. 334 (4): 1092–101.
5. Verma S, et al. (2003). Circulation 108 (6): 736–40.
6. Wellen KE, Hotamisligil GS (2005). J. Clin. Invest. 115 (5): 1111–9.
7. Lu SC, et al. (2002). FEBS Lett. 530 (1–3): 158–62.