Nori Mouse PLA2R ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of cPLA2R in mouse. 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 PLA2R has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PLA2R present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for PLA2R 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 PLA2R bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for cLA2R: Secretory phospholipase A2 receptor, PLA2R1
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Mouse PLA2R ELISA Kit Summary
Alternative names for PLA2R: Secretory phospholipase A2 receptor, PLA2R1,
| 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 | Q62028 |
| 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 | 30 pg/mL |
| Detection Range | 0.156-10 ng/mL |
| Specificity | Mouse PLA2R |
| 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:
Secretory phospholipase A2 receptor (PLA2R) is an enzyme that is encoded by the PLA2R1 gene.[1][2] M-type phospholipase A2 receptor is the major antigen in idiopathic membranous nephropathy attributed to over 70% of cases.[3] PLA2R interacts with PLA2 to release arachidonic acid from spleen cells.[4] circulating level of PLA2R increased during murine endotoxic shock.[5] PLA2R1 is increased in the airway epithelium in asthma, and serves as a regulator of airway hyperresponsiveness, airway permeability, antigen sensitization, and airway inflammation.[6] PLA2R likely exists as both a transmembrane form and a soluble form. The transmembrane receptor may play a role in clearance of phospholipase A2, thereby inhibiting its action. Polymorphisms of the gene have been associated with susceptibility to idiopathic membranous nephropathy.
References
- Ancian P, et al. (1995). The Journal of Biological Chemistry. 270 (15): 8963–70.
- Higashino K, et al. (1994). European Journal of Biochemistry / FEBS. 225 (1): 375–82.
- Beck LH, et al. (2009). The New England Journal of Medicine. 361 (1): 11–21.
- Morioka Y, et al. (2000). Archives of Biochemistry and Biophysics. 381 (1): 31–42.
- Yokota Y, et al. (2000). Archives of Biochemistry and Biophysics. 379 (1): 7–17.
- Nolin JD, et al. (2016) Am. J. Respir. Cell Mol. Biol. 55 (6), 825-836.
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