Nori Human ALOX5 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of ALOX5 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 ALOX5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ALOX5 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for ALOX5 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 ALOX5 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for ALOX5: Arachidonate 5-lipoxygenase, 5-lipoxygenase, 5-LOX, 5-LO, LOG5
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 ALOX5 ELISA Kit Summary
Alternative names for ALOX5: Arachidonate 5-lipoxygenase, 5-lipoxygenase, 5-LOX, 5-LO. LOG5
| 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 | P09917 |
| 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 | 3 pg/mL |
| Detection Range | 0.156-10 ng/mL |
| Specificity | Human ALOX5 |
| 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:
Arachidonate 5-lipoxygenase ( ALOX5, 5-lipoxygenase, 5-LOX, or 5-LO) is a non-heme iron-containing enzyme that is encoded by the ALOX5 gene.[1] ALOX5 is a member of the lipoxygenase family. It transforms essential fatty acids substrates into leukotrienes as well as a wide range of other biologically active products. ALOX5 is a current target for pharmaceutical intervention in a number of diseases. Cells primarily involved in regulating inflammation, allergy, and other immune responses, express ALOX5. However, platelets, T cells, and erythrocytes are ALOX5-negative. In skin, Langerhans cells strongly express ALOX5. Fibroblasts, smooth muscle cells and endothelial cells express low levels of ALOX5.[2] Aberrant expression of LOX5 is seen in various types of human cancer tumors in vivo as well as in various types of human cancer cell lines in vitro. ALOX5 products, particularly 5-hydroxyeicosatetraenoic acid and 5-oxo-eicosatetraenoic acid, promote the proliferation of these ALOX5 aberrantly expressing tumor cell lines suggesting that ALOX5 acts as a pro-malignancy factor for them and by extension their parent tumors.[6] Studies implicate ALOX5 in contributing to innate immunity by contributing to the mounting inflammatory responses to a wide range of acute and chronic diseases. These dual functions probably reflect ALOX5’s ability to form the: a) potent chemotactic factor, LTB4, and possibly also weaker chemotactic factor, 5S-HETE, which serve to attract and otherwise activate inflammation-inducing cells and b) lipoxin and resolvin subfamily of SPMs which tend to inhibit these cells as well as the overall inflammatory responses.[3][4]
References
- Funk CD, et al. (1989). Proc National Acad Sci of USA. 86 (8): 2587–91.
- Ochs MJ, et al. (2014). Basic & Clinical Pharmacology & Toxicology. 114 (1): 78–82.
- Rossi AG, O’Flaherty JT (1991). Lipids. 26 (12): 1184–8.
- Basil MC, Levy BD (2016). Nature Reviews. Immunology. 16 (1): 51–67.
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