Description
Human TNNC1 (Troponin C Type 1) ELISA Kit
The Human TNNC1 (Troponin C Type 1) ELISA Kit is specifically designed for the accurate measurement of TNNC1 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and consistent results for a variety of research purposes.Troponin C Type 1 is a vital regulator protein in muscle contraction, particularly in cardiac muscle. Elevated levels of TNNC1 in the bloodstream can indicate cardiac muscle damage, making it a valuable biomarker for diagnosing and monitoring conditions such as heart attacks and heart failure.
Researchers and healthcare professionals can rely on the Human TNNC1 ELISA Kit to facilitate their studies on cardiac muscle function and develop new approaches for managing cardiovascular diseases effectively.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 78.13-5000 pg/mL |
| Sensitivity: | 46.88 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human TNNC1 in samples. No significant cross-reactivity or interference between Human TNNC1 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 Human TNNC1. 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 Human TNNC1 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 TNNC1, 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 Human TNNC1. The concentration of Human TNNC1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | TNNC1: Troponin is the central regulatory protein of striated muscle contraction. Tn consists of three components: Tn-I which is the inhibitor of actomyosin ATPase, Tn-T which contains the binding site for tropomyosin and Tn-C. The binding of calcium to Tn-C abolishes the inhibitory action of Tn on actin filaments. Defects in TNNC1 are the cause of cardiomyopathy dilated type 1Z (CMD1Z). Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death. Defects in TNNC1 are the cause of familial hypertrophic cardiomyopathy type 13 (CMH13). A hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death. Belongs to the troponin C family. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Calcium-binding Chromosomal Location of Human Ortholog: 3p21. 1 Cellular Component: nucleoplasm; troponin complex; mitochondrion; cytosol; actin cytoskeleton Molecular Function:actin filament binding; troponin T binding; protein binding; protein homodimerization activity; troponin I binding; calcium ion binding; calcium-dependent protein binding Biological Process: response to metal ion; regulation of ATPase activity; regulation of muscle contraction; diaphragm contraction; ventricular cardiac muscle morphogenesis; regulation of muscle filament sliding speed; muscle filament sliding; cardiac muscle contraction Disease: Cardiomyopathy, Dilated, 1z; Cardiomyopathy, Familial Hypertrophic, 13 |
| NCBI Summary: | Troponin is a central regulatory protein of striated muscle contraction, and together with tropomyosin, is located on the actin filament. Troponin consists of 3 subunits: TnI, which is the inhibitor of actomyosin ATPase; TnT, which contains the binding site for tropomyosin; and TnC, the protein encoded by this gene. The binding of calcium to TnC abolishes the inhibitory action of TnI, thus allowing the interaction of actin with myosin, the hydrolysis of ATP, and the generation of tension. Mutations in this gene are associated with cardiomyopathy dilated type 1Z. [provided by RefSeq, Oct 2008] |
| UniProt Code: | Q6FH91 |
| NCBI GenInfo Identifier: | 49456725 |
| NCBI Gene ID: | 7134 |
| NCBI Accession: | CAG46683. 1 |
| UniProt Related Accession: | P63316 |
| Molecular Weight: | 18,403 Da |
| NCBI Full Name: | TNNC1, partial |
| NCBI Synonym Full Names: | troponin C type 1 (slow) |
| NCBI Official Symbol: | TNNC1 |
| NCBI Official Synonym Symbols: | TNC; TN-C; TNNC; CMD1Z; CMH13 |
| NCBI Protein Information: | troponin C, slow skeletal and cardiac muscles; cardiac troponin C; slow twitch skeletal/cardiac muscle troponin C; troponin C1, slow |
| UniProt Protein Name: | TNNC1 protein |
| UniProt Synonym Protein Names: | TNNC1 protein |
| Protein Family: | Troponin |
| UniProt Gene Name: | TNNC1 |
| UniProt Entry Name: | Q6FH91_HUMAN |
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 |
| 5000 | 2.557 2.605 | 2.581 | 2.529 |
| 2500 | 1.685 1.733 | 1.709 | 1.657 |
| 1250 | 1 0.964 | 0.982 | 0.93 |
| 625 | 0.448 0.474 | 0.461 | 0.409 |
| 312.5 | 0.257 0.247 | 0.252 | 0.2 |
| 156.25 | 0.166 0.158 | 0.162 | 0.11 |
| 78.13 | 0.106 0.11 | 0.108 | 0.056 |
| 0 | 0.044 0.06 | 0.052 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level HumanTNNC1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level HumanTNNC1 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) | 272.41 | 489.92 | 1860.99 | 274.16 | 538.09 | 1981.45 |
| Standard deviation | 16.18 | 23.86 | 67.93 | 17.16 | 27.82 | 68.16 |
| C V (%) | 5.94 | 4.87 | 3.65 | 6.26 | 5.17 | 3.44 |
Recovery
The recovery of HumanTNNC1 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) | 85-97 | 92 |
| EDTA plasma (n=5) | 86-98 | 93 |
| Cell culture media (n=5) | 88-102 | 95 |
Linearity
Samples were spiked with high concentrations of HumanTNNC1 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 (%) | 86-100 | 88-103 | 91-105 |
| Average (%) | 93 | 95 | 96 | |
| 1:4 | Range (%) | 90-104 | 83-96 | 82-96 |
| Average (%) | 96 | 90 | 88 | |
| 1:8 | Range (%) | 89-100 | 80-92 | 86-97 |
| Average (%) | 94 | 85 | 91 | |
| 1:16 | Range (%) | 90-106 | 84-97 | 87-99 |
| Average (%) | 97 | 90 | 92 |
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.
