Nori Rat Sirtuin 2 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of sirtuin 2 in rat. 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 SIRT2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any SIRT2 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for SIRT2 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 SIRT2 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for sirtuin 2: SIRT2, NAD-dependent protein deacetylase sirtuin-2,
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Rat Sirtuin 2 ELISA Kit Summary
Alternative names for sirtuin 2: SIRT2, NAD-dependent protein deacetylase sirtuin-2,
| 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 |
Q5RJQ4 |
| 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 | Rat Sirtuin 2 |
| 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:
Sirtuin proteins are a class of proteins that possess either mono-ADP-ribosyltransferase, or deacylase activity, including deacetylase, desuccinylase, demalonylase, demyristoylase and depalmitoylase activity.[1] Sirtuins regulate important biological pathways in bacteria, archaea and eukaryotes. The first sirtuin was identified in yeast and named sir2. Mammals possess seven sirtuins (SIRT1-7) that occupy different subcellular compartments such as the nucleus (SIRT1, -2, -6, -7), cytoplasm (SIRT1 and SIRT2) and the mitochondria (SIRT3, -4 and -5). Sirtuin activity is inhibited by nicotinamide, which binds to a specific receptor site,[2] so it is thought that drugs that interfere with this binding should increase sirtuin activity. Development of new agents that would specifically block the nicotinamide-binding site could provide an avenue for development of newer agents to treat degenerative diseases such as cancer, diabetes, atherosclerosis, and gout.[3] SIRT2 assists in the repair of DNA and regulates genes that undergo altered expression with age.[4] Adding resveratrol to the diet of mice inhibit gene expression profiles associated with muscle aging and age-related cardiac dysfunction.[5] A study performed on transgenic mice overexpressing SIRT6, showed an increased lifespan of about 15% in males. The transgenic males displayed lower serum levels of insulin-like growth factor 1 (IGF1) and changes in its metabolism, which may have contributed to the increased lifespan.[25]
References
- Yamamoto H, et al. (2007). Endocrinol. 21 (8): 1745–55.
- Avalos JL, et al. (2005). Cell 17 (6): 855–68.
- Taylor DM, et al. (2008). Mol. Life Sci. 65 (24): 4000–18.
- Oberdoerffer P, et al. (2008). Cell 135 (5): 907–18.
- Barger JL, et al. (2008). PLoS ONE 3 (6): e2264.
- Kanfi, Yariv et al. (2012). Nature 483 (7388): 218–21.
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