Description
Mouse PPAR- alpha (Peroxisome Proliferators-activator Receptors alpha) ELISA Kit
The Mouse PPAR Alpha (Peroxisome Proliferators Activator Receptors Alpha) ELISA Kit is designed for the precise and accurate measurement of PPAR Alpha levels in mouse serum, plasma, and cell culture supernatants. This kit is characterized by its high sensitivity and specificity, ensuring dependable and consistent results, making it an excellent tool for a variety of research purposes.PPAR Alpha is a key nuclear receptor involved in lipid metabolism, inflammation, and glucose homeostasis. It plays a critical role in various metabolic disorders, including obesity, diabetes, and cardiovascular diseases.
Understanding the levels of PPAR Alpha can provide valuable insights into these conditions and aid in the development of potential therapeutic strategies.Overall, the Mouse PPAR Alpha ELISA Kit offers a reliable and efficient method for quantifying PPAR Alpha levels in mouse samples, making it an indispensable tool for researchers studying metabolic disorders and related diseases.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.10 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse PPAR- alpha in samples. No significant cross-reactivity or interference between Mouse PPAR- alpha and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse PPAR- alpha. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse PPAR- alpha 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 Mouse PPAR- alpha, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse PPAR- alpha. The concentration of Mouse PPAR- alpha in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | PPAR-alpha: Ligand-activated transcription factor. Key regulator of lipid metabolism. Activated by the endogenous ligand 1-palmitoyl- 2-oleoyl-sn-glycerol-3-phosphocholine (16:0/18:1-GPC). Activated by oleylethanolamide, a naturally occurring lipid that regulates satiety. Receptor for peroxisome proliferators such as hypolipidemic drugs and fatty acids. Regulates the peroxisomal beta-oxidation pathway of fatty acids. Functions as transcription activator for the ACOX1 and P450 genes. Transactivation activity requires heterodimerization with RXRA and is antagonized by NR2C2. Belongs to the nuclear hormone receptor family. NR1 subfamily. |
| UniProt Protein Details: | Protein type:DNA-binding; Nuclear receptor Cellular Component: nucleus Molecular Function:protein domain specific binding; ligand-dependent nuclear receptor activity; NFAT protein binding; zinc ion binding; metal ion binding; receptor activity; drug binding; phosphatase binding; protein binding; DNA binding; ubiquitin conjugating enzyme binding; sequence-specific DNA binding; protein heterodimerization activity; protein complex binding; steroid hormone receptor activity; lipid binding; transcription factor activity Biological Process: transcription from RNA polymerase II promoter; epidermis development; wound healing; positive regulation of transcription, DNA-dependent; rhythmic process; behavioral response to nicotine; negative regulation of transcription from RNA polymerase II promoter; fatty acid metabolic process; negative regulation of appetite; response to insulin stimulus; positive regulation of gluconeogenesis; regulation of transcription, DNA-dependent; regulation of fatty acid metabolic process; negative regulation of blood pressure; circadian regulation of gene expression; negative regulation of protein binding; intracellular receptor-mediated signaling pathway; transcription, DNA-dependent; glucose metabolic process; lipoprotein metabolic process; regulation of circadian rhythm; regulation of gene expression; positive regulation of fatty acid oxidation; response to hypoxia; steroid hormone mediated signaling; positive regulation of transcription from RNA polymerase II promoter; lipid metabolic process |
| UniProt Code: | P23204 |
| NCBI GenInfo Identifier: | 1709728 |
| NCBI Gene ID: | 19013 |
| NCBI Accession: | P23204. 2 |
| UniProt Related Accession: | P23204 |
| Molecular Weight: | 52,347 Da |
| NCBI Full Name: | Peroxisome proliferator-activated receptor alpha |
| NCBI Synonym Full Names: | peroxisome proliferator activated receptor alpha |
| NCBI Official Symbol: | Ppara |
| NCBI Official Synonym Symbols: | Ppar; Nr1c1; AW742785; PPARalpha; PPAR-alpha; 4933429D07Rik |
| NCBI Protein Information: | peroxisome proliferator-activated receptor alpha; nuclear receptor subfamily 1 group C member 1 |
| UniProt Protein Name: | Peroxisome proliferator-activated receptor alpha |
| UniProt Synonym Protein Names: | Nuclear receptor subfamily 1 group C member 1 |
| Protein Family: | Peroxisome proliferator-activated receptor |
| UniProt Gene Name: | Ppara |
| UniProt Entry Name: | PPARA_MOUSE |
As the OD 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) | O.D | Average | Corrected |
| 10 | 2.463 2.475 | 2.469 | 2.401 |
| 5 | 1.549 1.581 | 1.565 | 1.497 |
| 2.5 | 0.929 0.919 | 0.924 | 0.856 |
| 1.25 | 0.425 0.433 | 0.429 | 0.361 |
| 0.63 | 0.258 0.24 | 0.249 | 0.181 |
| 0.32 | 0.167 0.167 | 0.167 | 0.099 |
| 0.16 | 0.118 0.12 | 0.119 | 0.051 |
| 0 | 0.061 0.075 | 0.068 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse PPAR- alpha were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse PPAR- alpha 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 | 1.14 | 3.99 | 0.51 | 1.11 | 4.08 |
| Standard deviation | 0.03 | 0.05 | 0.17 | 0.03 | 0.07 | 0.16 |
| C V (%) | 6.25 | 4.39 | 4.26 | 5.88 | 6.31 | 3.92 |
Recovery
The recovery of Mouse PPAR- alpha 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) | 91-106 | 99 |
| EDTA plasma (n=5) | 86-98 | 92 |
| Cell culture media (n=5) | 88-101 | 94 |
Linearity
Samples were spiked with high concentrations of Mouse PPAR- alpha 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 (%) | 86-100 | 98-115 | 90-105 |
| Average (%) | 93 | 105 | 97 | |
| 1:4 | Range (%) | 95-107 | 87-100 | 87-102 |
| Average (%) | 100 | 93 | 94 | |
| 1:8 | Range (%) | 92-103 | 86-101 | 84-97 |
| Average (%) | 98 | 93 | 90 | |
| 1:16 | Range (%) | 87-102 | 80-91 | 83-98 |
| Average (%) | 94 | 85 | 90 |
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 ELISA 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 | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| 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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
