Mouse Cell Death ELISA Kits
Mouse OPG (Osteoprotegerin) CLIA Kit (MOES00062)
- SKU:
- MOES00062
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 9.38pg/mL
- Range:
- 15.63-1000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- TNFRSF11B , OCIF, TR1
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Death
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection method: | Chemiluminescence |
| Detection range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Mouse OPG in samples. No significant cross-reactivity or interference between Mouse OPG 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 Mouse OPG. 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 Mouse OPG 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 OPG, 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 Mouse OPG. The concentration of Mouse OPG in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | TNFRSF11B: Acts as decoy receptor for RANKL and thereby neutralizes its function in osteoclastogenesis. Inhibits the activation of osteoclasts and promotes osteoclast apoptosis in vitro. Bone homeostasis seems to depend on the local RANKL/OPG ratio. May also play a role in preventing arterial calcification. May act as decoy receptor for TRAIL and protect against apoptosis. TRAIL binding blocks the inhibition of osteoclastogenesis. Defects in TNFRSF11B are the cause of juvenile Paget disease (JPD); also known as hyperostosis corticalis deformans juvenilis or hereditary hyperphosphatasia or chronic congenital idiopathic hyperphosphatasia. JPD is a rare autosomal recessive osteopathy that presents in infancy or early childhood. The disorder is characterized by rapidly remodeling woven bone, osteopenia, debilitating fractures, and deformities due to a markedly accelerated rate of bone remodeling throughout the skeleton. Approximately 40 cases of JPD have been reported worldwide. Unless it is treated with drugs that block osteoclast- mediated skeletal resorption, the disease can be fatal. |
| UniProt Protein Details: | Protein type:Secreted; Inhibitor; Secreted, signal peptide Cellular Component: proteinaceous extracellular matrix; extracellular space; extracellular region Biological Process: extracellular matrix organization and biogenesis; apoptosis; negative regulation of odontogenesis of dentine-containing teeth; negative regulation of bone resorption; signal transduction |
| UniProt Code: | O08712 |
| NCBI GenInfo Identifier: | 21264085 |
| NCBI Gene ID: | 18383 |
| NCBI Accession: | O08712. 1 |
| UniProt Secondary Accession: | O08712,O70202, |
| UniProt Related Accession: | O08712 |
| Molecular Weight: | 45,923 Da |
| NCBI Full Name: | Tumor necrosis factor receptor superfamily member 11B |
| NCBI Synonym Full Names: | tumor necrosis factor receptor superfamily, member 11b (osteoprotegerin) |
| NCBI Official Symbol: | Tnfrsf11b |
| NCBI Official Synonym Symbols: | Opg; TR1; OCIF |
| NCBI Protein Information: | tumor necrosis factor receptor superfamily member 11B; osteoprotegerin; osteoclastogenesis inhibitory factor |
| UniProt Protein Name: | Tumor necrosis factor receptor superfamily member 11B |
| UniProt Synonym Protein Names: | Osteoclastogenesis inhibitory factor; Osteoprotegerin |
| Protein Family: | Tumor necrosis factor receptor superfamily |
| UniProt Gene Name: | Tnfrsf11b |
| UniProt Entry Name: | TR11B_MOUSE |
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 (pg/mL) | RLU | Average | Corrected |
| 1000 | 53528 53822 | 53675 | 53650 |
| 500 | 22358 22620 | 22489 | 22464 |
| 250 | 10634 9788 | 10211 | 10186 |
| 125 | 4466 5336 | 4901 | 4876 |
| 62.5 | 2487 2419 | 2453 | 2428 |
| 31.25 | 1357 1205 | 1281 | 1256 |
| 15.63 | 678 738 | 708 | 683 |
| 0 | 24 26 | 25 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse OPG were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse OPG 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 (pg/mL) | 50.32 | 132.49 | 393.44 | 50.60 | 120.87 | 389.44 |
| Standard deviation | 5.50 | 13.92 | 24.12 | 4.82 | 12.24 | 30.92 |
| C V (%) | 10.93 | 10.51 | 6.13 | 9.53 | 10.13 | 7.94 |
Recovery
The recovery of Mouse OPG 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) | 97-112 | 105 |
| EDTA plasma (n=5) | 84-96 | 91 |
| Cell culture media (n=5) | 90-105 | 97 |
Linearity
Samples were spiked with high concentrations of Mouse OPG 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 (%) | 101-115 | 96-110 | 89-100 |
| Average (%) | 106 | 103 | 94 | |
| 1:4 | Range (%) | 100-117 | 99-115 | 102-116 |
| Average (%) | 107 | 107 | 109 | |
| 1:8 | Range (%) | 101-117 | 90-106 | 95-111 |
| Average (%) | 110 | 97 | 101 | |
| 1:16 | Range (%) | 87-102 | 103-117 | 87-98 |
| Average (%) | 93 | 110 | 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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