Nori Hyaluronan ELISA Kit
$461.00 – $832.00
DataSheet CoA SDS
This ELISA kit is for quantification of hyaluronan. 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 hyaluronan has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any hyaluronan present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for hyaluronan 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 hyaluronan bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for hyaluronan: Hyaluronic acid
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Hyaluronan ELISA Kit Summary
Alternative names for hyaluronan: Hyaluronic acid,
| 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 | na |
| 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 | hyaluronan |
| 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:
Hyaluronic acid (HA; conjugate base hyaluronate), also called hyaluronan, is an anionic, nonsulfated glycosaminoglycan distributed widely throughout connective, epithelial, and neural tissues. It is unique among glycosaminoglycans as it is non-sulfated, forms in the plasma membrane instead of the Golgi apparatus, and can be very large: human synovial HA averages about 7 million Da per molecule, or about 20,000 disaccharide monomers,[1] while other sources mention 3–4 million Da.[2] The molecular weight (size) of hyaluronan in cartilage decreases with age, but the amount increases.[3] The average 70 kg (150 lb) person has roughly 15 grams of hyaluronan in the body, one-third of which is turned over (i.e., degraded and synthesized) per day.[4] As one of the chief components of the extracellular matrix, it contributes significantly to cell proliferation and migration, and is involved in the progression of many malignant tumors.[5] Hyaluronic acid is also a component of the group A streptococcal. For example, hyaluronic acid is a major component of the synovial fluid and was found to increase the viscosity of the fluid.
Hyaluronic acid is an important component of articular cartilage, where it is present as a coat around each cell (chondrocyte). Hyaluronic acid is also a major component of skin, where it is involved in repairing tissue.
Hyaluronan also contributes to tissue hydrodynamics, movement, and proliferation of cells and participates in cell surface receptor interactions. Hyaluronan’s contribution to tumor growth may be due to its interaction with CD44. hyaluronan degradation products transduce their inflammatory signal through TLR2 and TLR4 in macrophages and dendritic cells. Hyaluronic acid has a key role in tissue regeneration, inflammation response, and angiogenesis, which are phases of skin wound repair.[6]
References
- Fraser JR, et al. (1997). J. Intern. Med. 242(1): 27–33.
- Saari H, et al. (1993). Inflammation. 17(4): 403–15.
- Holmes MW, et al. (1988). Biochem. J. 250(2): 435–441.
- Stern R (2004). Eur. J. Cell Biol. 83(7): 317–25.
- Itano, Naoki (2002). Proc Natl Acad Sci USA. 99(6): 3609–3614.
- Shaharudin, A.; Aziz, Z. (2016). Journal of Wound Care. 25(10): 585–592.
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