Nori Equine cPLA2 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of cPLA2 in equine. 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 cPLA2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any cPLA2 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for cPLA2 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 cPLA2 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for cPLA2: PLA2G4A, Phospholipase A2 G4A, cytosolic phospholipase A2, phospholipase A2 group IVA
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Equine cPLA2 ELISA Kit Summary
Alternative names for cPLA2: PLA2G4A, Phospholipase A2 G4A, cytosolic phospholipase A2, phospholipase A2 group IVA
Alternative names for equine: Horse
| 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 | O77793 |
| 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 | 25 pg/mL |
| Detection Range | 125-8000 pg/mL |
| Specificity | Equine cPLA2 |
| 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:
Phospholipase A2 G4A (PLA2G4A) is a member of the cytosolic phospholipase A2 group IV family. The enzyme catalyzes the hydrolysis of membrane phospholipids to release arachidonic acid which is subsequently metabolized into eicosanoids. Eicosanoids, including prostaglandins and leukotrienes, are lipid-based cellular hormones that regulate hemodynamics, inflammatory responses, and other intracellular pathways. The hydrolysis reaction also produces lysophospholipids that are converted into platelet-activating factor. The enzyme is activated by increased intracellular Ca2+ levels and phosphorylation, resulting in its translocation from the cytosol and nucleus to perinuclear membrane vesicles.[1] PLA2G4A has been shown to interact with HTATIP.[2] Mutations in this gene have been associated with multifocal stenosing ulceration of the small intestine.[2] Phospholipase activity is linked to energy metabolism,revealing cPLA2 as a central regulator of both lipidomics and energy flux.[3] Serine 228 is essential for catalytic activities of 85-kDa cytosolic phospholipase A2.[4] cPLA2 is phosphorylated and activated by MAP kinase.[5] A novel arachidonic acid-selective cytosolic PLA2 contains a Ca(2+)-dependent translocation domain with homology to PKC and GAP.[6]
References
- Sheridan AM, et al. (2001). Molecular and Cellular Biology. 21 (14): 4470–4481.
- Brooke MA, et al. (2012). Gut. 63: 96–104.
3. Slatter DA, et al. (2016) Cell Metab. 23 (5), 930-944.
4. Sharp JD,et al. (1994)J. Biol. Chem. 269 (37), 23250-23254.
5. Lin LL,et al. (1993) Cell 72 (2), 269-278.
6. Clark JD, et al. (1991) Cell 65 (6), 1043-1051.
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