Nori Feline Fetuin A ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of AHSG in cat. 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 AHSG has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any AHSG present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for AHSG 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 AHSG bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for fetuin A: alpha-2-HS-glycoprotein (AHSG)
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Feline Fetuin A ELISA Kit Summary
Alternative names for fetuin A: alpha-2-HS-glycoprotein (AHSG)
Alternative name for feline: cat
| 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 | A0A337SD37 |
| 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 | 600 pg/mL |
| Detection Range | 3.125-200 ng/mL |
| Specificity | Feline Fetuin A |
| 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:
Fetuin-A also known as alpha-2-HS-glycoprotein (AHSG) is a protein that in Felines is encoded by the AHSG gene.[1] Fetuin-A belongs to the fetuin class of plasma binding proteins and is more abundant in fetal than adult blood. It is synthesized by hepatocytes and is a secreted partially phosphorylated glycoprotein with complex proeolytic processing that circulates in blood and extracellular fluids. The AHSG molecule consists of two polypeptide chains, which are both cleaved from a proprotein encoded from a single mRNA. It is involved in several functions, such as endocytosis, brain development and the formation of bone tissue. The protein is commonly present in the cortical plate of the immature cerebral cortex and bone marrow hemopoietic matrix, and it has therefore been postulated that it participates in the development of the tissues. However, its exact significance is still obscure. The choroid plexus is an established extrahepatic expression site. Multiple post-translational modifications have been reported.[2] Fetuins are carrier proteins like albumin. Fetuin-A forms soluble complexes with calcium and phosphate and thus is a carrier of otherwise insoluble calcium phosphate.[3][4] Thus fetuin-A is a potent inhibitor of pathological calcification, in particular Calciphylaxis. Mice deficient in fetuin-A show systemic calcification of soft tissues.[5][6]
References
- Osawa M, et al. (1997). Gene. 196 (1–2): 121–5.
- Jahnen-Dechent W, et al. (1994). Eur. J. Biochem. 226 (1): 59–69.
- Heiss A, et al. (2008). J. Biol. Chem. 283 (21): 14815–25.
- Jahnen-Dechent W, et al. (2008). J. Mol. Med. 86 (4): 379–89.
- Westenfeld R, et al. (2007). Nephrol. Dial. Transplant. 22 (6): 1537–46.
- Schafer C, et al. (2003). J. Clin. Invest. 112 (3): 357–66.
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