Description
Human CPN10 (Chaperonin 10) ELISA Kit
The Human CPN10 (Chaperonin-10) ELISA Kit is specifically designed for the accurate quantification of CPN10 levels in human samples including serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and consistent results for a variety of research purposes.CPN10, also known as Chaperonin-10, is a critical molecular chaperone involved in protein folding and cellular stress response. It plays a crucial role in maintaining protein homeostasis and protecting cells from damage.
Dysregulation of CPN10 has been linked to various diseases and disorders, highlighting its importance as a potential biomarker for disease diagnosis and monitoring.With the Human CPN10 ELISA Kit, researchers can accurately measure CPN10 levels in biological samples, enabling detailed investigations into the role of CPN10 in physiological and pathological processes. This kit is an essential tool for studying molecular chaperones, cellular stress responses, and their implications in health and disease.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.10 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human CPN10 in samples. No significant cross-reactivity or interference between Human CPN10 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 CPN10. 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 CPN10 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 CPN10, 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 CPN10. The concentration of Human CPN10 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | HSPE1: Eukaryotic CPN10 homolog which is essential for mitochondrial protein biogenesis, together with CPN60. Binds to CPN60 in the presence of Mg-ATP and suppresses the ATPase activity of the latter. Belongs to the GroES chaperonin family. |
| UniProt Protein Details: | Protein type:Mitochondrial Chromosomal Location of Human Ortholog: 2q33. 1 Cellular Component: membrane; mitochondrial matrix; mitochondrion Molecular Function:ATP binding; chaperone binding; protein binding; unfolded protein binding Biological Process: caspase activation; osteoblast differentiation; protein folding; response to unfolded protein |
| NCBI Summary: | This gene encodes a major heat shock protein which functions as a chaperonin. Its structure consists of a heptameric ring which binds to another heat shock protein in order to form a symmetric, functional heterodimer which enhances protein folding in an ATP-dependent manner. This gene and its co-chaperonin, HSPD1, are arranged in a head-to-head orientation on chromosome 2. Naturally occurring read-through transcription occurs between this locus and the neighboring locus MOBKL3. [provided by RefSeq, Feb 2011] |
| UniProt Code: | P61604 |
| NCBI GenInfo Identifier: | 47606335 |
| NCBI Gene ID: | 3336 |
| NCBI Accession: | P61604. 2 |
| UniProt Secondary Accession: | P61604,O95421, Q04984, Q53X54, Q9UDH0, |
| UniProt Related Accession: | P61604 |
| Molecular Weight: | 10,932 Da |
| NCBI Full Name: | 10 kDa heat shock protein, mitochondrial |
| NCBI Synonym Full Names: | heat shock protein family E (Hsp10) member 1 |
| NCBI Official Symbol: | HSPE1 |
| NCBI Official Synonym Symbols: | EPF; CPN10; GROES; HSP10 |
| NCBI Protein Information: | 10 kDa heat shock protein, mitochondrial |
| UniProt Protein Name: | 10 kDa heat shock protein, mitochondrial |
| UniProt Synonym Protein Names: | 10 kDa chaperonin; Chaperonin 10; CPN10; Early-pregnancy factor; EPF |
| UniProt Gene Name: | HSPE1 |
| UniProt Entry Name: | CH10_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 (ng/mL) | O.D | Average | Corrected |
| 10 | 2.478 2.49 | 2.484 | 2.43 |
| 5 | 1.6 1.652 | 1.626 | 1.572 |
| 2.5 | 0.908 0.902 | 0.905 | 0.851 |
| 1.25 | 0.442 0.45 | 0.446 | 0.392 |
| 0.63 | 0.238 0.22 | 0.229 | 0.175 |
| 0.31 | 0.163 0.141 | 0.152 | 0.098 |
| 0.16 | 0.1 0.112 | 0.106 | 0.052 |
| 0 | 0.051 0.057 | 0.054 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CPN10 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CPN10 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 (ng/mL) | 0.50 | 1.20 | 4.80 | 0.50 | 1.20 | 4.70 |
| Standard deviation | 0.03 | 0.06 | 0.19 | 0.03 | 0.06 | 0.19 |
| C V (%) | 6.00 | 5.00 | 3.96 | 6.00 | 5.00 | 4.04 |
Recovery
The recovery of Human CPN10 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) | 95-110 | 100 |
| EDTA plasma (n=5) | 86-100 | 94 |
| Cell culture media (n=5) | 88-103 | 95 |
Linearity
Samples were spiked with high concentrations of Human CPN10 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 (%) | 89-102 | 92-102 | 86-99 |
| Average (%) | 94 | 97 | 93 | |
| 1:4 | Range (%) | 101-115 | 82-94 | 90-105 |
| Average (%) | 107 | 88 | 97 | |
| 1:8 | Range (%) | 94-107 | 84-95 | 93-107 |
| Average (%) | 101 | 90 | 100 | |
| 1:16 | Range (%) | 96-110 | 85-96 | 95-113 |
| Average (%) | 102 | 91 | 103 |
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.
