Description
Human CX43 (Connexin 43) ELISA Kit
The Human CX43 (Connexin 43) ELISA Kit is specially designed for the precise measurement of Connexin 43 levels in human serum, plasma, and cell culture supernatants. This ELISA kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results for a variety of research purposes.Connexin 43 is a key protein that plays a vital role in cell-to-cell communication, facilitating the transfer of ions and small molecules between cells. Dysregulation of Connexin 43 has been linked to various health conditions, including cardiac diseases, skin disorders, and neurological disorders.
Therefore, this ELISA kit serves as a valuable tool for studying Connexin 43 function and potential therapeutic interventions for related diseases.Don't miss out on the opportunity to enhance your research with the Human CX43 ELISA Kit from Assay Genie. Visit the website for more information and to order your kit today!
| 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 CX43 in samples. No significant cross-reactivity or interference between Human CX43 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 CX43. 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 CX43 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 CX43, 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 CX43. The concentration of Human CX43 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | GJA1: an integral membrane protein of the connexin family, alpha-type (group II) subfamily. Hexamers of connexin-43 form connexons, which aggregate together to form gap junctions, through which materials of low MW diffuse from one cell to a neighboring cell. May play a critical role in the physiology of hearing by participating in the recycling of potassium to the cochlear endolymph. |
| UniProt Protein Details: | Protein type:Membrane protein, integral; Channel, misc. ; Motility/polarity/chemotaxis; Membrane protein, multi-pass Chromosomal Location of Human Ortholog: 6q22. 31 Cellular Component: Golgi apparatus; endoplasmic reticulum membrane; focal adhesion; contractile fiber; integral to plasma membrane; lysosome; early endosome; intermediate filament; fascia adherens; cytosol; lipid raft; Golgi membrane; multivesicular body; connexon complex; mitochondrial outer membrane; apical plasma membrane; gap junction; plasma membrane; lateral plasma membrane Molecular Function:protein binding; signal transducer activity; ion transmembrane transporter activity; beta-tubulin binding; SH3 domain binding; gap junction channel activity; receptor binding; PDZ domain binding Biological Process: lens development in camera-type eye; response to peptide hormone stimulus; apoptosis; heart development; neuron migration; milk ejection; positive regulation of vasodilation; signal transduction; elevation of cytosolic calcium ion concentration; muscle contraction; cell-cell signaling; negative regulation of cardiac muscle cell proliferation; positive regulation of glomerular filtration; transport; response to glucose stimulus; positive regulation of striated muscle development; heart looping; ATP transport; adult heart development; chronic inflammatory response; positive regulation of I-kappaB kinase/NF-kappaB cascade; gap junction assembly; in utero embryonic development; epithelial cell maturation; regulation of bone remodeling; positive regulation of insulin secretion; skeletal muscle regeneration; regulation of calcium ion transport; vascular transport; protein oligomerization; osteoblast differentiation; positive regulation of osteoblast differentiation; negative regulation of endothelial cell proliferation; positive regulation of vasoconstriction; positive regulation of protein catabolic process; blood vessel morphogenesis; embryonic digit morphogenesis; regulation of bone mineralization; neurite morphogenesis; response to pH Disease: Syndactyly, Type Iii; Craniometaphyseal Dysplasia, Autosomal Recessive; Palmoplantar Keratoderma And Congenital Alopecia 1; Atrioventricular Septal Defect 3; Oculodentodigital Dysplasia; Erythrokeratodermia Variabilis Et Progressiva; Oculodentodigital Dysplasia, Autosomal Recessive; Hypoplastic Left Heart Syndrome 1 |
| NCBI Summary: | This gene is a member of the connexin gene family. The encoded protein is a component of gap junctions, which are composed of arrays of intercellular channels that provide a route for the diffusion of low molecular weight materials from cell to cell. The encoded protein is the major protein of gap junctions in the heart that are thought to have a crucial role in the synchronized contraction of the heart and in embryonic development. A related intronless pseudogene has been mapped to chromosome 5. Mutations in this gene have been associated with oculodentodigital dysplasia, autosomal recessive craniometaphyseal dysplasia and heart malformations. [provided by RefSeq, May 2014] |
| UniProt Code: | P17302 |
| NCBI GenInfo Identifier: | 117706 |
| NCBI Gene ID: | 2697 |
| NCBI Accession: | P17302. 2 |
| UniProt Secondary Accession: | P17302,Q6FHU1, Q9Y5I8, B2R5U9, |
| UniProt Related Accession: | P17302 |
| Molecular Weight: | 382 |
| NCBI Full Name: | Gap junction alpha-1 protein |
| NCBI Synonym Full Names: | gap junction protein, alpha 1, 43kDa |
| NCBI Official Symbol: | GJA1 |
| NCBI Official Synonym Symbols: | HSS; CMDR; CX43; GJAL; ODDD; AVSD3; HLHS1 |
| NCBI Protein Information: | gap junction alpha-1 protein; connexin 43; connexin-43; gap junction 43 kDa heart protein |
| UniProt Protein Name: | Gap junction alpha-1 protein |
| UniProt Synonym Protein Names: | Connexin-43; Cx43; Gap junction 43 kDa heart protein |
| Protein Family: | Gap junction alpha-1 protein |
| UniProt Gene Name: | GJA1 |
| UniProt Entry Name: | CXA1_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 | 2.577 2.635 | 2.606 | 2.518 |
| 5 | 1.729 1.729 | 1.729 | 1.641 |
| 2.5 | 1.075 1.055 | 1.065 | 0.977 |
| 1.25 | 0.47 0.486 | 0.478 | 0.39 |
| 0.63 | 0.312 0.288 | 0.3 | 0.212 |
| 0.32 | 0.204 0.194 | 0.199 | 0.111 |
| 0.16 | 0.141 0.147 | 0.144 | 0.056 |
| 0 | 0.081 0.095 | 0.088 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CX43 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CX43 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) | 0.54 | 1.53 | 3.36 | 0.54 | 1.56 | 3.43 |
| Standard deviation | 0.04 | 0.08 | 0.13 | 0.03 | 0.08 | 0.16 |
| C V (%) | 7.41 | 5.23 | 3.87 | 5.56 | 5.13 | 4.66 |
Recovery
The recovery of Human CX43 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) | 94-107 | 100 |
| EDTA plasma (n=5) | 88-100 | 93 |
| Cell culture media (n=5) | 93-105 | 99 |
Linearity
Samples were spiked with high concentrations of Human CX43 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 (%) | 99-113 | 98-111 | 85-99 |
| Average (%) | 106 | 105 | 91 | |
| 1:4 | Range (%) | 87-99 | 85-98 | 83-94 |
| Average (%) | 93 | 90 | 88 | |
| 1:8 | Range (%) | 89-103 | 83-94 | 88-100 |
| Average (%) | 94 | 89 | 94 | |
| 1:16 | Range (%) | 90-102 | 80-95 | 83-96 |
| Average (%) | 95 | 87 | 89 |
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.
