Protein Overview
CD3E encodes the CD3 epsilon chain, a signal-transducing subunit of the TCR-CD3 complex that pairs with CD3 delta and CD3 gamma alongside the TCR heterodimer and CD3 zeta homodimer. Its ITAM motifs are phosphorylated by LCK/FYN upon TCR engagement, recruiting ZAP70 and initiating T cell activation; CD3E is also essential for thymic T cell development.
Expected Localization
Plasma membrane, assembled as part of the multi-subunit TCR-CD3 complex.
Produces uniform membranous/surface staining on T lymphocytes (pan-T-cell marker); some intracellular/perinuclear signal may appear during complex assembly in the ER/Golgi.
Expected Expression Pattern
Pan-T-cell marker expressed on virtually all TCRαβ+ and TCRγδ+ T lymphocytes and thymocytes; found in blood, lymph node, tonsil, spleen, and thymus. Not expressed on B cells, mature NK cells, or myeloid cells; surface density varies with TCR internalization following activation.
Relevant Pathways
- TCR-CD3 complex assembly and surface expression
- ITAM phosphorylation by LCK/FYN
- ZAP70 recruitment and activation
- Downstream MAPK/ERK, PLCγ1/Ca²⁺, and NF-κB/NFAT signaling
- T cell development and thymic selection
Clone Information
Antibody Species: Rabbit
Cell Type Compatibility: Validated in human cells only using Cytos™ kits.
For more details on the clone and reactivity with other species, visit ABclonal and search for the clone catalog # A19017BCS.
Qualitative Results
Localization Assessment
CD3E protein panel demonstrates strong localization to the CD3-positive cell membrane. The observed localization of CD3E protein is consistent with published data.
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| Jurkat | Raji |
Jurkat and Raji cells were cultured independently in flasks, spun down to a 12-well format following the user guide for suspension cells, fixed with 4% formaldehyde for 30 minutes, and permeabilized using 70% ethanol for 10 minutes. Cells were then incubated with probe made with biotinylated Rabbit monoclonal anti-CD3E antibody for 1 hour at room temperature. The slide kit was then assembled into a flow cell and loaded onto an AVITI24™ along with the Cytos Protein Cartridge for a multiomics cell screening run. CD3E demonstrated expected expression in Jurkat and no obvious expression in Raji cells. These results support high confidence in target-specific detection and validate this antibody as a passing clone.
Quantitative Results
| Signal-to-noise ratio Acceptance criteria: > 2x background |
Pass |
- 24.45 ± 2.39 (Jurkat; n=2 runs)
Indicates the fold brightness of the target signal as measured by the Mean Intensity Above (Otsu-defined) Threshold (IAT), over the background (a channel-matched isotype control) in the same cell. The result of 10.37 in the CD3-positive Jurkat line is over 5 times the acceptance threshold — a clean, easily distinguishable signal in the positive condition.
|
Panel-level run-to-run reproducibility |
Pass |
For each imaging panel, we assess reproducibility by comparing the intensity signal a given antibody target produces across independent replicate runs. Within each run, we measure the average per-well fluorescence intensity for every target in the panel. We then plot each target's intensity from one run against the matching target's intensity from another run of the same layout, and calculate the Pearson correlation coefficient (r) between those two sets of measurements. Squaring that value gives the R² (coefficient of determination), which reports what fraction of the variation in one run's signal is explained by the other run's signal.
Disclaimer: This information is provided to help assess whether the target suits your experiment. It reflects internal testing conditions and does not guarantee performance in your application. Performance in other cell types and species such as mouse or rat may require additional validation using Cytos kits as performance is not guaranteed.
ELEMENT BIOSCIENCES For Research Use Only. Not for Use in diagnostic procedures.