Nori Human ANGPTL3 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of ANGPTL3 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 ANGPTL3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ANGPTL3 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for ANGPTL3 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 ANGPTL3 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for ANTPGL3: Angiopoietin-like 3, ANGPT5
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 ANGPTL3 ELISA Kit Summary
Alternative names for ANGPTL3: Angiopoietin-like 3, ANGPT5
| 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 Antigen Accession Number |
Q9Y5C1 |
| 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 | 25 pg/mL |
| Detection Range | 125-8000 pg/mL |
| Specificity | Human ANGPTL3 |
| 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:
Angiopoietin-like 3 ( ANGPTL3) is a protein that is encoded by the ANGPTL3 gene[1] and a member of the angiopoietin-like family of secreted factors. It is expressed predominantly in the liver, and has the characteristic structure of angiopoietins, consisting of a signal peptide, N-terminal coiled-coil domain, and the C-terminal fibrinogen (FBN)-like domain. The FBN-like domain in ANGPTL3 protein was shown to bind alpha-5/beta-3 integrins, and this binding induced endothelial cell adhesion and migration. ANGPTL3 may also play a role in the regulation of angiogenesis. Angptl3 also acts as dual inhibitor of lipoprotein lipase (LPL) and endothelial lipase (EL),[2] thereby increasing plasma triglyceride, LDL cholesterol and HDL cholesterol in mice and humans.[2] ANGPTL3 inhibits endothelial lipase hydrolysis of HDL-phospholipid (PL), thereby increasing HDL-PL levels. Circulating PL-rich HDL particles have high cholesterol efflux abilities. Angptl3 plays a major role in promoting uptake of circulating triglycerides into white adipose tissue in the fed state,[3] likely through activation by Angptl8, a feeding-induced hepatokine,[4][5] to inhibit postprandial LPL activity in cardiac and skeletal muscles,[6] as suggested by the ANGPTL3-4-8 model.[7] ANGPTL3 is a determinant factor of HDL level and positively correlates with plasma HDL cholesterol. In humans with genetic loss-of-function variants in one copy of ANGPTL3, the serum LDL-C levels are reduced. In those with loss-of-function variants in both copies of ANGPTL3, low LDL-C, low HDL-C, and low triglycerides are seen (“familial combined hypolipidemia“).[8]
References
- Conklin D, et al. (1999). Genomics. 62 (3): 477–82.
- Tikka A, Jauhiainen M (2016). Endocrine. 52 (2): 187–93.
- Wang Y, et al. (2015). Proc Natl Acad Sci USA. 112 (37): 11630–5.
- Zhang R (2012). Biochem Biophys Res Communications. 424 (4): 786–92.
- Ren G, et al. (2012). Am J Physiol Endocrinology and Metabolism. 303 (3): E334-51.
- Fu Z, et al. (2015). Scientific Reports. 5: 18502. doi:1038/srep18502.
- Zhang R (2016). Open Biology. 6 (4): 150272. doi:1098/rsob.150272.
- Musunuru K, et al. (2010). The New England Journal of Medicine. 363 (23): 2220–7.
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