Nori Human Hyaluronidase-1 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of hyaluronidase-1 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 HYAL1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HYAL1 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for HYAL1 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 HYAL1 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for hyaluronidase-1: HYAL1, LUCA1
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 Hyaluronidase-1 ELISA Kit Summary
Alternative names for hyaluronidase-1: HYAL1, LUCA1
| 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 | Q12794 |
| 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 | Human HYAL1 |
| 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:
Hyaluronidase-1 is an enzyme that is encoded by the HYAL1 gene.[1] The enzyme is 435 amino acids long with a molecular weight of 55-60 kDa.[2] HYAL1 is composed of two closely associated domains: a N-terminal catalytic domain (Phe22-Thr352) and a smaller C-terminal domain (Ser353-Trp435).[3] Hyaluronidases intracellularly degrade hyaluronan, one of the major glycosaminoglycans of the extracellular matrix. HYA1 is active at an acidic pH and is the major hyaluronidase in plasma. Mutations in this gene are associated with mucopolysaccharidosis type IX, or hyaluronidase deficiency. HYAL1 is responsible for the hydrolysis of intracellular hyaluronan of all sizes into fragments as small as tetrasaccharides.[3] HYAL1 is implicated in several types of cancers, likely due to the angiogenic effects of HYAL1-cleaved hyaluronan fragments. [4] In bladder, prostate, and head and neck carcinomas, elevated hyaluronan and HYAL1 levels are found in tumor cells, tissues, and related body fluids (e.g. urine for bladder cancer and saliva for head and neck cancer). Urinary hyaluronan and hyaluronidase levels, measured by the HA-HAase test, have ~88% accuracy in detecting bladder cancer, regardless of the tumor grade and stage.[5] In breast cancer, HYAL1 is also overexpressed in cell lines MDA-MB-231 and MCF-7 and invasive duct cancer tissues and metastatic lymph nodes.[6] Higher HYAL1 expression has also been detected in primary tumor tissue from patients with subsequent brain metastases versus those without.[7]
References
- Frost GI, et al. (1997). Biochem Biophysical Res Communications. 236 (1): 10–5.
- Tan JX, et al. (2011). PLOS ONE. 6 (7) e22836. doi:10.1371/journal.pone.0022836.
- Chao KL, et al. (2007). Biochemistry. 46 (23): 6911–20.
- Slevin M, et al. (1998). Lab Investigation; J Tech Methods Pathol. 78 (8): 987–1003.
- Lokeshwar VB, et al. (2000). The Journal of Urology. 163 (1): 348–56.
- Tan JX, Wang et al. (2011). International Journal of Cancer. 128 (6): 1303–15.
- Witzel I, et al. (2017). Breast Cancer Research and Treatment. 162 (3): 427–438.
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