Nori Human Desnutrin ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of desnutrin in human. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for ATGL has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ATGL present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for ATGL is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of ATGL bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for desnutrin: adipose triglyceride lipase (ATGL), patatin-like phospholipase domain-containing protein 2 (PNPLA2)
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Human Desnutrin ELISA Kit Summary
Alternative names for desnutrin: adipose triglyceride lipase (ATGL), patatin-like phospholipase domain-containing protein 2 (PNPLA2)
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antiben Accession Number | Q96AD5 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 30 pg/mL |
| Detection Range | 0.156-10 ng/mL |
| Specificity | Human desnutrin |
| 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 |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
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:
Desnutrin also known as adipose triglyceride lipase (ATGL), patatin-like phospholipase domain-containing protein 2 (PNPLA2), is an enzyme that is encoded by the PNPLA2 gene.[1] Desnutrin, an adipocyte gene encoding a novel patatin domain-containing protein, is induced by fasting and glucocorticoids and ectopic expression of desnutrin increases triglyceride hydrolysis.[2] Fat mobilization in adipose tissue is promoted by adipose triglyceride lipase.[3] ATGL has a key role in lipid droplet/adiposome degradation in mammalian cells.[4] The ATGL gene is associated with free fatty acids, triglycerides, and type 2 diabetes.[5] Adipose triglyceride lipase-mediated lipolysis of cellular fat stores is activated by CGI-58 and defective in Chanarin-Dorfman Syndrome.[6] Human adipose triglyceride lipase (PNPLA2) is not regulated by obesity and exhibits low in vitro triglyceride hydrolase activity.[7] Adipose triglyceride lipase and hormone-sensitive lipase protein expression is decreased in the obese insulin-resistant state.[8]
References
- Wilson PA, et al. (2006). J Lipid Res. 47 (9): 1940–9.
- Villena JA, et al. (2004). J. Biol. Chem. 279 (45): 47066–75.
- Zimmermann R, et al. (2004). Science. 306 (5700): 1383–6.
- Smirnova E, et al. (2006). EMBO Rep. 7 (1): 106–13.
- Schoenborn V, et al. (2006). Diabetes. 55 (5): 1270–5.
- Lass A, et al. (2007Cell Metab. 3 (5): 309–19.
- Mairal A, et al. (2007). Diabetologia. 49 (7): 1629–36.
- Jocken JW, et al. (2007). J. Clin. Endocrinol. Metab. 92 (6): 2292–9.
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