Nori Mouse Visfatin ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of visfatin in mouse. 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 NAMPT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any NAMPT present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for NAMPT 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 NAMPT bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for visfatin: Extracellular nicotinamide phosphoribosyl transferase, eNAMPT, NAMPT
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Mouse Visfatin ELISA Kit Summary
Alternative names for visfatin: Extracellular nicotinamide phosphoribosyl transferase, eNAMPT, NAMPT
| 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 |
Q99KQ4 |
| 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 | 300 pg/mL |
| Detection Range | 1.56-100 ng/mL |
| Specificity | Mouse visfatin |
| 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:
Visfatin is an extracellular form of nicotinamide phosphoribosyl transferase (eNAMPT) and is encoded by the NAMPT gene.[1] Visfatin is functionally different from intracellular NAMPT (iNAMPT).[2] The sirtuin 1 is required for eNAMPT secretion from adipose tissue.[3] eNAMPT may act more as a cytokine, although its receptor (possibly TLR4) has not been proven.[4] It has been demonstrated that eNAMPT could bind to and activate TLR4.[5] eNAMPT can exist as a dimer or monomer, but is normally a circulating dimer.[3] As a monomer, eNAMPT has pro-inflammatory effects that are independent of NAD+, whereas the dimeric form of eNAMPT protects against these effects.[3] visfatin was originally cloned as a putative cytokine shown to enhance the maturation of B cell precursors in the presence of Interleukin-7 (IL-7) and stem cell factor, it was therefore named “pre-B cell colony-enhancing factor” (PBEF).[1] Visfatin is a dimeric type II phosphoribosyl transferase enzyme involved in NAD biosynthesis.[6][7] eNAMPT has been shown to be more enzymatically active than iNAMPT, supporting the proposal that eNAMPT from adipose tissue enhances NAD+ in tissues with low levels of iNAMPT, notably pancreatic beta cells and brain neurons.[8]
References
- Samal B, et al. (1994). Molecular and Cellular Biology. 14 (2): 1431–7.
- Grolla AA, et al. (2016). British Journal of Pharmacology. 173 (14): 2182–2194.
- Yoshino J, et al. (2018). Cell Metabolism. 27 (3): 513–528.
- Galli U, et al. (2020). Frontiers in Pharmacology. 11: 656.
- Garcia, Joe G. N.; et al. (2015). Scientific Reports. 5: 13135.
- Wang T, et al. (2006). Nature Structural & Molecular Biology. 13 (7): 661–2. doi:1038/nsmb1114.
- Kim MK, et al. (2006). Journal of Molecular Biology. 362 (1): 66–77.
- Imai S (2016). npj Systems Biology and Applications. 2: 16018.
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