Mouse Cardiovascular ELISA Kits
Mouse TM (Thrombomodulin) CLIA Kit (MOES00559)
- SKU:
- MOES00559
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 9.38pg/mL
- Range:
- 15.63-1000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- THBD, AHUS6, BDCA3, CD141, THPH12, THRM
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cardiovascular
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 TM in samples. No significant cross-reactivity or interference between Mouse TM 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 TM. 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 TM 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 TM, 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 TM. The concentration of Mouse TM in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | thrombomodulin: Thrombomodulin is a specific endothelial cell receptor that forms a 1:1 stoichiometric complex with thrombin. This complex is responsible for the conversion of protein C to the activated protein C (protein Ca). Once evolved, protein Ca scissions the activated cofactors of the coagulation mechanism, factor Va and factor VIIIa, and thereby reduces the amount of thrombin generated. Defects in THBD are the cause of thrombophilia due to thrombomodulin defect (THPH12). A hemostatic disorder characterized by a tendency to thrombosis. Defects in THBD are a cause of susceptibility to hemolytic uremic syndrome atypical type 6 (AHUS6). An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype. |
| UniProt Protein Details: | Protein type:Membrane protein, integral Cellular Component: extracellular space; cell surface; membrane; integral to plasma membrane; plasma membrane; integral to membrane Molecular Function:transmembrane receptor activity; calcium ion binding Biological Process: embryonic development; hemostasis; negative regulation of blood coagulation; female pregnancy; blood coagulation; negative regulation of coagulation |
| UniProt Code: | P15306 |
| NCBI GenInfo Identifier: | 136171 |
| NCBI Gene ID: | 21824 |
| NCBI Accession: | P15306. 1 |
| UniProt Related Accession: | P15306 |
| Molecular Weight: | |
| NCBI Full Name: | Thrombomodulin |
| NCBI Synonym Full Names: | thrombomodulin |
| NCBI Official Symbol: | Thbd |
| NCBI Official Synonym Symbols: | TM; CD141; AI385582 |
| NCBI Protein Information: | thrombomodulin |
| UniProt Protein Name: | Thrombomodulin |
| UniProt Synonym Protein Names: | Fetomodulin; CD_antigen: CD141 |
| Protein Family: | Thrombomodulin |
| UniProt Gene Name: | Thbd |
| UniProt Entry Name: | TRBM_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 | 52821 54529 | 53675 | 53650 |
| 500 | 21958 23020 | 22489 | 22464 |
| 250 | 10683 9739 | 10211 | 10186 |
| 125 | 4699 5103 | 4901 | 4876 |
| 62.5 | 2621 2285 | 2453 | 2428 |
| 31.25 | 1281 1281 | 1281 | 1256 |
| 15.63 | 664 752 | 708 | 683 |
| 0 | 25 25 | 25 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse TM were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse TM 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) | 48.18 | 156.77 | 423.85 | 47.65 | 159.35 | 462.91 |
| Standard deviation | 5.32 | 12.54 | 47.85 | 4.07 | 13.10 | 28.84 |
| C V (%) | 11.04 | 8.00 | 11.29 | 8.54 | 8.22 | 6.23 |
Recovery
The recovery of Mouse TM 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) | 100-115 | 107 |
| EDTA plasma (n=5) | 100-116 | 107 |
| Cell culture media (n=5) | 97-110 | 104 |
Linearity
Samples were spiked with high concentrations of Mouse TM 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 (%) | 87-99 | 88-100 | 92-104 |
| Average (%) | 93 | 95 | 98 | |
| 1:4 | Range (%) | 100-112 | 104-118 | 85-98 |
| Average (%) | 106 | 110 | 91 | |
| 1:8 | Range (%) | 93-106 | 103-115 | 95-109 |
| Average (%) | 99 | 109 | 101 | |
| 1:16 | Range (%) | 93-108 | 91-107 | 101-115 |
| Average (%) | 100 | 97 | 109 |
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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