Mouse Cell Biology ELISA Kits 2
Mouse EMILIN1 (Elastin Microfibril Interface Located Protein 1) CLIA Kit (MOES00238)
- SKU:
- MOES00238
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 4.69pg/mL
- Range:
- 7.81-500pg/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection method: | Chemiluminescence |
| Detection range: | 7.81-500 pg/mL |
| Sensitivity: | 4.69 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Mouse EMILIN1 in samples. No significant cross-reactivity or interference between Mouse EMILIN1 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 EMILIN1. 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 EMILIN1 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 EMILIN1, 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 EMILIN1. The concentration of Mouse EMILIN1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | Function: May be responsible for anchoring smooth muscle cells to elastic fibers, and may be involved not only in the formation of the elastic fiber, but also in the processes that regulate vessel assembly. Has cell adhesive capacity |
| UniProt Protein Details: | By similarity. May have a function in placenta formation and initial organogenesis and a later role in interstitial connective tissue. Subunit structure: Homotrimer associated through a moderately stable interaction of the C-terminal globular C1q domains, allowing the nucleation of the triple helix and then a further quaternary assembly to higher-order polymers via intermolecular disulfide bonds. Interacts with EMILIN2 By similarity. Subcellular location: Secreted extracellular space extracellular matrix. Note: Found mainly at the interface between amorphous elastin and microfibrils. Developmental stage: Detectable in morula and blastocyst. First expressed in ectoplacental cone in embryos of 6. 5 days and in extraembryonic visceral endoderm at 7. 5 days. Expressed also in the allantois. Expression in the ectoplacental cone-derived secondary trophoblast giant cells and spongiotrophoblast is strong up to 11. 5 days and then declines. In the embryo, high levels are initially expressed in blood vessels, perineural mesenchyme and somites at 8. 5 days. Later on, intense expression is identified in the mesenchymal component of organs anlage (ie lung and liver) and different mesenchymal condensations (ie limb bud and branchial arches). At late gestation expression is widely distributed in interstitial connective tissue and smooth muscle cell-rich tissues. Ref. 3 Sequence similarities: Contains 1 C1q domain. Contains 1 collagen-like domain. Contains 1 EMI domain. |
| UniProt Code: | Q99K41 |
| NCBI GenInfo Identifier: | 19527130 |
| NCBI Gene ID: | 100952 |
| NCBI Accession: | NP_598679. 1 |
| UniProt Related Accession: | Q99K41 |
| Molecular Weight: | |
| NCBI Full Name: | EMILIN-1 |
| NCBI Synonym Full Names: | elastin microfibril interfacer 1 |
| NCBI Official Symbol: | Emilin1 |
| NCBI Official Synonym Symbols: | gp115; AW229038; BB105748; EMILIN-1; 5830419M17Rik |
| NCBI Protein Information: | EMILIN-1; elastin microfibril interface located protein 1; elastin microfibril interface-located protein 1 |
| UniProt Protein Name: | EMILIN-1 |
| UniProt Synonym Protein Names: | Elastin microfibril interface-located protein 1 |
| Protein Family: | EMILIN |
| UniProt Gene Name: | Emilin1 |
| UniProt Entry Name: | EMIL1_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 |
| 500 | 50463 52611 | 51537 | 51508 |
| 250 | 19264 22732 | 20998 | 20969 |
| 125 | 10128 8664 | 9396 | 9367 |
| 62.5 | 4194 4830 | 4512 | 4483 |
| 31.25 | 2368 2230 | 2299 | 2270 |
| 15.63 | 1341 1159 | 1250 | 1221 |
| 7.81 | 689 791 | 740 | 711 |
| 0 | 28 30 | 29 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse EMILIN1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse EMILIN1 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) | 25.72 | 54.22 | 235.36 | 27.59 | 51.96 | 216.55 |
| Standard deviation | 2.55 | 5.81 | 17.79 | 3.38 | 5.78 | 16.98 |
| C V (%) | 9.91 | 10.72 | 7.56 | 12.25 | 11.12 | 7.84 |
Recovery
The recovery of Mouse EMILIN1 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) | 96-110 | 103 |
| EDTA plasma (n=5) | 100-110 | 105 |
| Cell culture media (n=5) | 97-110 | 104 |
Linearity
Samples were spiked with high concentrations of Mouse EMILIN1 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 (%) | 101-117 | 101-116 | 92-108 |
| Average (%) | 109 | 108 | 99 | |
| 1:4 | Range (%) | 94-104 | 100-116 | 101-118 |
| Average (%) | 99 | 108 | 108 | |
| 1:8 | Range (%) | 97-109 | 99-115 | 90-105 |
| Average (%) | 103 | 107 | 98 | |
| 1:16 | Range (%) | 87-99 | 101-116 | 92-106 |
| Average (%) | 93 | 109 | 99 |
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.
Related Products
Rat EMILIN1 (Elastin Microfibril Interface Located Protein 1) ELISA Kit
