Description
Mouse tPA (Plasminogen Activator, Tissue) ELISA Kit
The Mouse TPA (Tissue Plasminogen Activator) ELISA Kit is a reliable tool for the precise measurement of TPA levels in mouse serum, plasma, and tissue samples. This kit offers exceptional sensitivity and specificity, guaranteeing consistent and accurate results for various research purposes.TPA is a key enzyme involved in the breakdown of blood clots, making it vital for maintaining proper blood flow and preventing clot-related complications. Dysregulation of TPA activity has been linked to various cardiovascular diseases, making it a valuable biomarker for studying these conditions and exploring potential treatment options.
With its user-friendly protocol and excellent performance, the Mouse TPA ELISA Kit is an indispensable tool for researchers investigating the role of TPA in health and disease. Get reliable data and valuable insights with this advanced ELISA kit from Assay Genie.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 39.06-2500 pg/mL |
| Sensitivity: | 23.44 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse tPA in samples. No significant cross-reactivity or interference between Mouse tPA 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 tPA. 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 tPA 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 tPA, 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 tPA. The concentration of Mouse tPA in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | PLAT: Converts the abundant, but inactive, zymogen plasminogen to plasmin by hydrolyzing a single Arg-Val bond in plasminogen. By controlling plasmin-mediated proteolysis, it plays an important role in tissue remodeling and degradation, in cell migration and many other physiopathological events. Plays a direct role in facilitating neuronal migration. Increased activity of TPA results in increased fibrinolysis of fibrin blood clots that is associated with excessive bleeding. Defective release of TPA results in hypofibrinolysis that can lead to thrombosis or embolism. Belongs to the peptidase S1 family. 4 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Secreted, signal peptide; Protease; EC 3. 4. 21. 68; Secreted; Vesicle; Cytoskeletal Cellular Component: extracellular space; cell surface; apical part of cell; cytoplasm; extracellular region; synapse; secretory granule Molecular Function:peptidase activity; hydrolase activity; serine-type peptidase activity; serine-type endopeptidase activity; catalytic activity Biological Process: negative regulation of proteolysis; synaptic transmission, glutamatergic; smooth muscle cell migration; response to hypoxia; platelet-derived growth factor receptor signaling pathway; proteolysis; plasminogen activation |
| NCBI Summary: | This gene encodes a key enzyme of the fibrinolytic pathway. The encoded protein undergoes proteolytic processing by plasmin to generate a heterodimeric serine protease that cleaves the proenzyme plasminogen to produce plasmin, a protease that is required to break down fibrin clots. Additionally, the encoded protein is involved in other biological processes such as synaptic plasticity, cell migration and tissue remodeling. Mice lacking the encoded protein display a reduction in long-term potentiation in hippocampus and conversely, transgenic mice overexpressing the encoded protein have increased and prolonged long-term potentiation. [provided by RefSeq, Jul 2015] |
| UniProt Code: | P11214 |
| NCBI GenInfo Identifier: | 170763477 |
| NCBI Gene ID: | 18791 |
| NCBI Accession: | NP_032898. 2 |
| UniProt Related Accession: | P11214 |
| Molecular Weight: | Predicted MW: 20. 7kDaAccurate MW: 22kDa |
| NCBI Full Name: | tissue-type plasminogen activator preproprotein |
| NCBI Synonym Full Names: | plasminogen activator, tissue |
| NCBI Official Symbol: | Plat |
| NCBI Official Synonym Symbols: | tPA; AU020998; AW212668; D8Ertd2e |
| NCBI Protein Information: | tissue-type plasminogen activator |
| UniProt Protein Name: | Tissue-type plasminogen activator |
| Protein Family: | Taurine--pyruvate aminotransferase |
| UniProt Gene Name: | Plat |
| UniProt Entry Name: | TPA_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 (pg/mL) | O.D | Average | Corrected |
| 2500 | 2.34 2.344 | 2.342 | 2.256 |
| 1250 | 1.465 1.501 | 1.483 | 1.397 |
| 625 | 0.849 0.827 | 0.838 | 0.752 |
| 312.5 | 0.456 0.49 | 0.473 | 0.387 |
| 156.25 | 0.267 0.245 | 0.256 | 0.17 |
| 78.13 | 0.175 0.173 | 0.174 | 0.088 |
| 39.06 | 0.126 0.138 | 0.132 | 0.046 |
| 0 | 0.078 0.094 | 0.086 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse tPA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse tPA 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) | 119.26 | 295.20 | 1060.03 | 115.55 | 308.64 | 1015.25 |
| Standard deviation | 7.86 | 13.46 | 35.19 | 5.84 | 16.36 | 55.23 |
| C V (%) | 6.59 | 4.56 | 3.32 | 5.05 | 5.30 | 5.44 |
Recovery
The recovery of Mouse tPA 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) | 88-101 | 95 |
| EDTA plasma (n=5) | 91-107 | 98 |
| Cell culture media (n=5) | 94-109 | 101 |
Linearity
Samples were spiked with high concentrations of Mouse tPA 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 (%) | 91-105 | 89-105 | 92-104 |
| Average (%) | 98 | 97 | 97 | |
| 1:4 | Range (%) | 87-103 | 83-96 | 84-97 |
| Average (%) | 94 | 88 | 89 | |
| 1:8 | Range (%) | 86-100 | 87-99 | 86-98 |
| Average (%) | 93 | 92 | 93 | |
| 1:16 | Range (%) | 90-104 | 84-96 | 85-96 |
| Average (%) | 96 | 90 | 91 |
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.
