Description
Human CT (Calcitonin) ELISA Kit
The Human CT (Calcitonin) ELISA Kit is a reliable and accurate tool for detecting levels of calcitonin in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for various research applications.Calcitonin is a hormone that helps regulate calcium levels in the body, primarily by inhibiting bone breakdown and lowering blood calcium levels.
Abnormal levels of calcitonin can indicate conditions such as hypercalcemia, osteoporosis, and certain types of cancer. The Human CT (Calcitonin) ELISA Kit is essential for studying these conditions and developing potential therapies.Overall, the Human CT (Calcitonin) ELISA Kit is a valuable tool for researchers and clinicians interested in studying calcitonin-related diseases and developing targeted treatments.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human CT in samples. No significant cross-reactivity or interference between Human CT 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 CT. 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 CT 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 CT, 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 CT. The concentration of Human CT in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | CALCA: Calcitonin causes a rapid but short-lived drop in the level of calcium and phosphate in blood by promoting the incorporation of those ions in the bones. Belongs to the calcitonin family. 3 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide Chromosomal Location of Human Ortholog: 11p15. 2 Cellular Component: extracellular region; extracellular space Molecular Function:calcitonin receptor binding; identical protein binding; protein binding Biological Process: activation of protein kinase activity; cellular protein metabolic process; elevation of cytosolic calcium ion concentration; elevation of cytosolic calcium ion concentration during G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); embryo implantation; monocyte chemotaxis; negative regulation of bone resorption; negative regulation of transcription, DNA-dependent; positive regulation of adenylate cyclase activity |
| NCBI Summary: | This gene encodes the peptide hormones calcitonin, calcitonin gene-related peptide and katacalcin by tissue-specific alternative RNA splicing of the gene transcripts and cleavage of inactive precursor proteins. Calcitonin is involved in calcium regulation and acts to regulate phosphorus metabolism. Calcitonin gene-related peptide functions as a vasodilator and as an antimicrobial peptide while katacalcin is a calcium-lowering peptide. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Aug 2014] |
| UniProt Code: | P01258 |
| NCBI GenInfo Identifier: | 322510018 |
| NCBI Gene ID: | 796 |
| NCBI Accession: | P01258. 2 |
| UniProt Secondary Accession: | P01258,Q13935, Q13937, Q52LX7, B7ZL39, |
| UniProt Related Accession: | P01258,P06881 |
| Molecular Weight: | 38. 6kDa |
| NCBI Full Name: | Calcitonin |
| NCBI Synonym Full Names: | calcitonin related polypeptide alpha |
| NCBI Official Symbol: | CALCA |
| NCBI Official Synonym Symbols: | CT; KC; PCT; CGRP; CALC1; CGRP1; CGRP-I |
| NCBI Protein Information: | calcitonin; calcitonin gene-related peptide 1 |
| UniProt Protein Name: | Calcitonin |
| UniProt Synonym Protein Names: | CalcitoninKatacalcinAlternative name(s):Calcitonin carboxyl-terminal peptide; CCP |
| UniProt Gene Name: | CALCA |
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 |
| 1000 | 2.419 2.477 | 2.448 | 2.369 |
| 500 | 1.564 1.594 | 1.579 | 1.5 |
| 250 | 0.998 0.996 | 0.997 | 0.918 |
| 125 | 0.43 0.468 | 0.449 | 0.37 |
| 62.5 | 0.294 0.266 | 0.28 | 0.201 |
| 31.25 | 0.188 0.172 | 0.18 | 0.101 |
| 15.63 | 0.127 0.135 | 0.131 | 0.052 |
| 0 | 0.073 0.085 | 0.079 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CT were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CT 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) | 46.50 | 119.00 | 450.80 | 43.80 | 109.60 | 473.30 |
| Standard deviation | 3.20 | 6.30 | 20.30 | 2.30 | 5.70 | 24.10 |
| C V (%) | 6.88 | 5.29 | 4.50 | 5.25 | 5.20 | 5.09 |
Recovery
The recovery of Human CT 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) | 89-99 | 94 |
| EDTA plasma (n=5) | 90-106 | 98 |
| Cell culture media (n=5) | 91-103 | 97 |
Linearity
Samples were spiked with high concentrations of Human CT 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 (%) | 98-114 | 92-106 | 99-114 |
| Average (%) | 104 | 99 | 104 | |
| 1:4 | Range (%) | 102-114 | 84-94 | 98-111 |
| Average (%) | 108 | 89 | 104 | |
| 1:8 | Range (%) | 92-106 | 84-95 | 93-106 |
| Average (%) | 100 | 90 | 100 | |
| 1:16 | Range (%) | 99-117 | 84-94 | 94-111 |
| Average (%) | 107 | 89 | 102 |
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 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 90 µL 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.
