Human CD3e (Cluster of Differentiation 3e) ELISA Kit (HUES03607)
- SKU:
- HUES03607
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P07766
- Sensitivity:
- 0.19ng/mL
- Range:
- 0.31-20ng/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Immunology
Description
Human CD3e (Cluster of Differentiation 3e) ELISA Kit
The Human CD3e (Cluster of Differentiation 3e) ELISA Kit is specifically designed for the accurate detection of CD3e levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring reliable and reproducible results for a variety of research applications.CD3e is a critical protein found on T cells, an essential component of the immune system responsible for recognizing and fighting off infections and diseases.
By measuring CD3e levels, researchers can gain valuable insights into the immune response and potentially diagnose and monitor immune-related disorders.Overall, the Human CD3e ELISA Kit from Assay Genie provides researchers with a powerful tool to study the role of CD3e in immune function and disease, ultimately contributing to the development of new therapeutic approaches.
Assay type: | Sandwich |
Format: | 96T |
Assay time: | 4.5h |
Reactivity: | Human |
Detection Method: | Colormetric |
Detection Range: | 0.31-20 ng/mL |
Sensitivity: | 0.19 ng/mL |
Sample Volume Required Per Well: | 100µL |
Sample Type: | Serum, plasma and other biological fluids |
Specificity: | This kit recognizes Human CD3e in samples. No significant cross-reactivity or interference between Human CD3e 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 CD3e. 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 CD3e 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 CD3e, 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 CD3e. The concentration of Human CD3e in samples can be calculated by comparing the OD of the samples to the standard curve.
UniProt Protein Function: | CD3E: a T cell surface glycoprotein that is a component of the T cell antigen receptor. The recruitment of Nck by CD3 epsilon reveals a ligand-induced conformational change essential for T cell receptor signaling and synapse formation. Contains 1 immunoglobulin-like domain and 1 ITAM domain. |
UniProt Protein Details: | Protein type:Receptor, misc. ; Membrane protein, integral Chromosomal Location of Human Ortholog: 11q23 Cellular Component: T cell receptor complex; integral to plasma membrane; plasma membrane; immunological synapse; alpha-beta T cell receptor complex; intercellular junction; external side of plasma membrane Molecular Function:protein binding; transmembrane receptor activity; receptor signaling complex scaffold activity; protein heterodimerization activity; receptor signaling protein activity; T cell receptor binding; SH3 domain binding; protein kinase binding Biological Process: regulation of immune response; T cell activation; positive regulation of interleukin-2 biosynthetic process; signal complex assembly; positive regulation of calcium-mediated signaling; negative thymic T cell selection; T cell receptor signaling pathway; positive regulation of interleukin-4 production; regulation of apoptosis; G-protein coupled receptor protein signaling pathway; positive regulation of interferon-gamma production; positive regulation of peptidyl-tyrosine phosphorylation; cell surface receptor linked signal transduction; negative regulation of smoothened signaling pathway; T cell costimulation; protein complex assembly; positive regulation of alpha-beta T cell proliferation; positive regulation of T cell proliferation; positive regulation of T cell anergy; transmembrane receptor protein tyrosine kinase signaling pathway; response to nutrient Disease: Immunodeficiency 18 |
NCBI Summary: | The protein encoded by this gene is the CD3-epsilon polypeptide, which together with CD3-gamma, -delta and -zeta, and the T-cell receptor alpha/beta and gamma/delta heterodimers, forms the T-cell receptor-CD3 complex. This complex plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. The genes encoding the epsilon, gamma and delta polypeptides are located in the same cluster on chromosome 11. The epsilon polypeptide plays an essential role in T-cell development. Defects in this gene cause immunodeficiency. This gene has also been linked to a susceptibility to type I diabetes in women. [provided by RefSeq, Jul 2008] |
UniProt Code: | P07766 |
NCBI GenInfo Identifier: | 1345708 |
NCBI Gene ID: | 916 |
NCBI Accession: | P07766. 2 |
UniProt Related Accession: | P07766 |
Molecular Weight: | |
NCBI Full Name: | T-cell surface glycoprotein CD3 epsilon chain |
NCBI Synonym Full Names: | CD3e molecule |
NCBI Official Symbol: | CD3E |
NCBI Official Synonym Symbols: | T3E; TCRE; IMD18 |
NCBI Protein Information: | T-cell surface glycoprotein CD3 epsilon chain |
UniProt Protein Name: | T-cell surface glycoprotein CD3 epsilon chain |
UniProt Synonym Protein Names: | T-cell surface antigen T3/Leu-4 epsilon chain; CD_antigen: CD3e |
UniProt Gene Name: | CD3E |
UniProt Entry Name: | CD3E_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 |
20 | 2.408 2.414 | 2.411 | 2.356 |
10 | 1.551 1.601 | 1.576 | 1.521 |
5 | 0.975 0.947 | 0.961 | 0.906 |
2.5 | 0.447 0.457 | 0.452 | 0.397 |
1.25 | 0.249 0.241 | 0.245 | 0.19 |
0.63 | 0.167 0.147 | 0.157 | 0.102 |
0.31 | 0.098 0.116 | 0.107 | 0.052 |
0 | 0.045 0.065 | 0.055 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CD3e were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CD3e 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) | 1.01 | 2.39 | 8.60 | 0.96 | 2.59 | 8.61 |
Standard deviation | 0.05 | 0.13 | 0.43 | 0.05 | 0.14 | 0.47 |
C V (%) | 4.95 | 5.44 | 5.00 | 5.21 | 5.41 | 5.46 |
Recovery
The recovery of Human CD3e 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) | 85-98 | 90 |
EDTA plasma (n=5) | 94-106 | 100 |
Cell culture media (n=5) | 84-98 | 91 |
Linearity
Samples were spiked with high concentrations of Human CD3e 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 (%) | 88-101 | 93-108 | 90-103 |
Average (%) | 93 | 100 | 97 | |
1:4 | Range (%) | 90-105 | 86-98 | 85-97 |
Average (%) | 97 | 91 | 90 | |
1:8 | Range (%) | 87-98 | 79-91 | 84-98 |
Average (%) | 93 | 85 | 90 | |
1:16 | Range (%) | 93-104 | 84-99 | 87-97 |
Average (%) | 98 | 91 | 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.