Human Developmental Biology ELISA Kits
Human GPR120 (G Protein Coupled Receptor 120) CLIA Kit (HUES00736)
- SKU:
- HUES00736
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 0.09ng/mL
- Range:
- 0.16-10ng/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Developmental Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 0.16-10 ng/mL |
| Sensitivity: | 0.09 ng/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human GPR120 in samples. No significant cross-reactivity or interference between Human GPR120 and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human GPR120. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human GPR120 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human GPR120, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human GPR120. The concentration of Human GPR120 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | GPR120: Receptor for medium and long-chain free fatty acid (FAA). Signals via a G(q)/G(11)-coupled pathway. Acts as a receptor for omega-3 fatty acids and mediates robust anti- inflammatory effects particularly in macrophages and fat cells. The anti-inflammatory effects involve inhibition of TAK1 through a beta-arrestin 2 (ARRB2)/TAB1 dependent effect but independent of G(q)/G(11)-coupled pathway. Mediates potent insulin sensitizing and antidiabetic effects by repressing macrophage-induced tissue inflammation. May mediate the taste of fatty acids. Belongs to the G-protein coupled receptor 1 family. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Lipid-binding; Receptor, GPCR; GPCR, family 1; Membrane protein, integral; Membrane protein, multi-pass Chromosomal Location of Human Ortholog: 10q23. 33 Cellular Component: integral to plasma membrane; plasma membrane Molecular Function:fatty acid binding; peptide binding; taste receptor activity Biological Process: cellular response to hormone stimulus; hormone secretion; negative regulation of apoptosis; negative regulation of cytokine secretion; negative regulation of inflammatory response Disease: Body Mass Index Quantitative Trait Locus 10 |
| NCBI Summary: | This gene encodes a G protein-coupled receptor (GPR) which belongs to the rhodopsin family of GPRs. The encoded protein functions as a receptor for free fatty acids, including omega-3, and participates in suppressing anti-inflammatory responses and insulin sensitizing. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Feb 2012] |
| UniProt Code: | Q5NUL3 |
| NCBI GenInfo Identifier: | 82581671 |
| NCBI Gene ID: | 338557 |
| NCBI Accession: | Q5NUL3. 2 |
| UniProt Secondary Accession: | Q5NUL3,Q495H1, Q5VY25, Q5VY26, Q7Z605, Q86SM7, |
| UniProt Related Accession: | Q5NUL3 |
| Molecular Weight: | 40,494 Da |
| NCBI Full Name: | Free fatty acid receptor 4 |
| NCBI Synonym Full Names: | free fatty acid receptor 4 |
| NCBI Official Symbol: | FFAR4 |
| NCBI Official Synonym Symbols: | GT01; PGR4; BMIQ10; GPR120; GPR129; O3FAR1 |
| NCBI Protein Information: | free fatty acid receptor 4 |
| UniProt Protein Name: | Free fatty acid receptor 4 |
| UniProt Synonym Protein Names: | G-protein coupled receptor 120; G-protein coupled receptor 129; G-protein coupled receptor GT01; G-protein coupled receptor PGR4; Omega-3 fatty acid receptor 1 |
| Protein Family: | Free fatty acid receptor |
| UniProt Gene Name: | FFAR4 |
| UniProt Entry Name: | FFAR4_HUMAN |
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (ng/mL) | RLU | Average | Corrected |
| 10 | 30536 34406 | 32471 | 32447 |
| 5 | 15411 15529 | 15470 | 15446 |
| 2.5 | 7729 7277 | 7503 | 7479 |
| 1.25 | 3638 3668 | 3653 | 3629 |
| 0.63 | 1872 1650 | 1761 | 1737 |
| 0.31 | 860 788 | 824 | 800 |
| 0.16 | 347 367 | 357 | 333 |
| 0 | 24 24 | 24 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GPR120 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GPR120 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 0.48 | 1.03 | 4.78 | 0.44 | 1.09 | 4.40 |
| Standard deviation | 0.06 | 0.11 | 0.37 | 0.05 | 0.09 | 0.31 |
| C V (%) | 12.50 | 10.68 | 7.74 | 11.36 | 8.26 | 7.05 |
Recovery
The recovery of Human GPR120 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 101-114 | 108 |
| EDTA plasma (n=5) | 98-110 | 105 |
| Cell culture media (n=5) | 91-107 | 98 |
Linearity
Samples were spiked with high concentrations of Human GPR120 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 98-111 | 100-117 | 97-111 |
| Average (%) | 103 | 107 | 104 | |
| 1:4 | Range (%) | 97-115 | 96-111 | 97-111 |
| Average (%) | 105 | 102 | 103 | |
| 1:8 | Range (%) | 95-107 | 99-114 | 97-108 |
| Average (%) | 101 | 108 | 103 | |
| 1:16 | Range (%) | 95-107 | 88-98 | 98-114 |
| Average (%) | 101 | 93 | 106 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro CLIA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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