Nori Porcine PNPLA3 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of PNPLA3 in porcine. 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 PNPLA3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PNPLA3 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for PNPLA3 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 PNPLA3 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names forPNPLA3: Patatin-like phospholipase domain-containing protein 3, adiponutrin, ADPN, acylglycerol O-acyltransferase, calcium-independent phospholipase A2-epsilon, iPLA2-epsilon
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Porcine PNPLA3 ELISA Kit Summary
Alternative names for PNPLA3: Patatin-like phospholipase domain-containing protein 3, adiponutrin, ADPN, acylglycerol O-acyltransferase, calcium-independent phospholipase A2-epsilon, iPLA2-epsilon
Alternative names for porcine: Pig, swine
| 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 | A5JUR7 |
| 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 | 120 pg/mL |
| Detection Range | 6.25-400 ng/mL |
| Specificity | Porcine PNPLA3 |
| 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:
Patatin-like phospholipase domain-containing protein 3 (PNPLA3) also known as adiponutrin (ADPN), acylglycerol O-acyltransferase or calcium-independent phospholipase A2-epsilon (iPLA2-epsilon) is an enzyme that is encoded by the PNPLA3 gene.[1][2][3] Adiponutrin is a triacylglycerol lipase that mediates triacylglycerol hydrolysis in adipocytes. The encoded protein, which appears to be membrane bound, may be involved in the balance of energy usage/storage in adipocytes. An association between alcoholic liver disease in caucasians and variations in this gene has been confirmed.[4] A mutation of isoleucine to methionine (I[ATC]>M[ATG]) SNP rs738409 has been confirmed to increase susceptibility to non-alcoholic liver disease and also to have effects in diabetes.[5]
References
- Wilson PA, et al. (2006). Journal of Lipid Research. 47 (9): 1940–9.
- Kienesberger PC, et al. (2009). Journal of Lipid Research. 50 Suppl: S63-8.
- Jenkins CM, et al. (2004). The Journal of Biological Chemistry. 279 (47): 48968–75.
- Stickel F, Hampe J (2012). Gut. 61 (1): 150–9.
- Pingitore P, Romeo S (2019). Biochimica et Biophysica Acta (BBA) – Molecular and Cell Biology of Lipids. 1864 (6): 900–906.
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