Human Metabolism ELISA Kits
Human ADIPOR2 (Adiponectin Receptor 2) CLIA Kit (HUES00203)
- SKU:
- HUES00203
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 0.09ng/mL
- Range:
- 0.16-10ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- ACDCR2, PAQR2
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Metabolism
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: | Tissue homogenates,cell lysates and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human ADIPOR2 in samples. No significant cross-reactivity or interference between Human ADIPOR2 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 ADIPOR2. 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 ADIPOR2 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 ADIPOR2, 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 ADIPOR2. The concentration of Human ADIPOR2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | ADIPOR2: Receptor for globular and full-length adiponectin (APM1), an essential hormone secreted by adipocytes that acts as an antidiabetic. Probably involved in metabolic pathways that regulate lipid metabolism such as fatty acid oxidation. Mediates increased AMPK, PPARA ligand activity, fatty acid oxidation and glucose uptake by adiponectin. Has some intermediate-affinity receptor activity for both globular and full-length adiponectin. Belongs to the ADIPOR family. |
| UniProt Protein Details: | Protein type:Membrane protein, integral; Membrane protein, multi-pass; Receptor, misc. Chromosomal Location of Human Ortholog: 12p13. 31 Cellular Component: integral to membrane; plasma membrane Molecular Function:adiponectin binding; identical protein binding; hormone binding; protein heterodimerization activity; receptor activity Biological Process: positive regulation of glucose import; hormone-mediated signaling; heart development; adiponectin-mediated signaling pathway; fatty acid oxidation; female pregnancy; negative regulation of cell growth; response to nutrient |
| NCBI Summary: | The adiponectin receptors, ADIPOR1 (MIM 607945) and ADIPOR2, serve as receptors for globular and full-length adiponectin (MIM 605441) and mediate increased AMPK (see MIM 602739) and PPAR-alpha (PPARA; MIM 170998) ligand activities, as well as fatty acid oxidation and glucose uptake by adiponectin (Yamauchi et al. , 2003 [PubMed 12802337]). [supplied by OMIM, Mar 2008] |
| UniProt Code: | Q86V24 |
| NCBI GenInfo Identifier: | 38372190 |
| NCBI Gene ID: | 79602 |
| NCBI Accession: | Q86V24. 1 |
| UniProt Related Accession: | Q86V24 |
| Molecular Weight: | 43kDa |
| NCBI Full Name: | Adiponectin receptor protein 2 |
| NCBI Synonym Full Names: | adiponectin receptor 2 |
| NCBI Official Symbol: | ADIPOR2 |
| NCBI Official Synonym Symbols: | PAQR2; ACDCR2 |
| NCBI Protein Information: | adiponectin receptor protein 2 |
| UniProt Protein Name: | Adiponectin receptor protein 2 |
| UniProt Synonym Protein Names: | Progestin and adipoQ receptor family member II |
| Protein Family: | Adiponectin receptor protein |
| UniProt Gene Name: | ADIPOR2 |
| UniProt Entry Name: | ADR2_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 | 32361 32581 | 32471 | 32447 |
| 5 | 14700 16240 | 15470 | 15446 |
| 2.5 | 7712 7294 | 7503 | 7479 |
| 1.25 | 3466 3840 | 3653 | 3629 |
| 0.63 | 1779 1743 | 1761 | 1737 |
| 0.31 | 831 817 | 824 | 800 |
| 0.16 | 339 375 | 357 | 333 |
| 0 | 24 24 | 24 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ADIPOR2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ADIPOR2 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) | 0.55 | 1.15 | 4.89 | 0.54 | 1.22 | 4.86 |
| Standard deviation | 0.04 | 0.10 | 0.52 | 0.05 | 0.10 | 0.40 |
| C V (%) | 7.27 | 8.70 | 10.63 | 9.26 | 8.20 | 8.23 |
Recovery
The recovery of Human ADIPOR2 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) | 95-111 | 101 |
| EDTA plasma (n=5) | 96-112 | 104 |
| Cell culture media (n=5) | 91-102 | 97 |
Linearity
Samples were spiked with high concentrations of Human ADIPOR2 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 (%) | 99-113 | 93-109 | 102-116 |
| Average (%) | 105 | 101 | 108 | |
| 1:4 | Range (%) | 95-109 | 99-111 | 99-118 |
| Average (%) | 101 | 104 | 107 | |
| 1:8 | Range (%) | 87-99 | 83-97 | 95-110 |
| Average (%) | 94 | 90 | 101 | |
| 1:16 | Range (%) | 92-106 | 91-105 | 88-101 |
| Average (%) | 97 | 97 | 93 |
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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