- NCBI Gene Id:
- Protein Name:
- T-cell surface glycoprotein CD3 epsilon chain
- Alias Symbols:
- T3E, TCRE, FLJ18683, CD3E
- Replacement Item:
- This antibody may replace item sc-1127 from Santa Cruz Biotechnology.
- Description of Target:
- MOUSE ANTI HUMAN CD3
- Protein Size (# AA):
- IHC-AFF, FC
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express CD3E.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express CD3E.
- The immunogen for anti-CD3E antibody: human infant thymocytes and lymphocytes from a patient with Sezary Syndrome
- Predicted Species Reactivity:
- Predicted Homology Based on Immunogen Sequence:
- Product Format:
- Purified IgG - liquid
- Reconstitution and Storage:
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Printable datasheet for anti-CD3E antibody - OASA01817
- Application Info:
- Flow Cytometry: 1/100 - 1/200
- Application Data:
- Application #1: Staining of human peripheral blood lymphocytes with Mouse anti Human CD3: FITC
Application #2: Staining of human peripheral blood lymphocytes with Mouse anti Human CD3
Application #3: Staining of human peripheral blood lymphocytes with Mouse anti Human CD3: RPE- Alexa Fluor 647
Application #4: Staining of human peripheral blood lymphocytes with Mouse anti Human CD3: RPE- Alexa Fluor 750
Application #5: Staining of human peripheral blood lymphocytes with Mouse anti Human CD3:Alexa Fluor 405
- Additional Information:
- Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells of the P3/NS1/1-Ag4-1 mouse myeloma cell line.
- Preparation: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Preservative Stabilisers: 0.09% - Sodium Azide
- Approx Protein Conc: IgG concentration 1 mg/ml
- Protocol Information:
- Citation: 1: Mahon NG, Madden BP, Caforio AL, Elliott PM, Haven AJ, Keogh BE, Davies MJ, McKenna WJ. Immunohistologic evidence of myocardial disease in apparently healthy relatives of patients with dilated cardiomyopathy. J Am Coll Cardiol. 2002 Feb 6;39(3):455-62. PubMed PMID: 11823084.
Experiment Name: Histology and immunohistochemistry
Experiment Background: Recent immunohistochemical studies have shown that patients with DCM have increased expression of human leukocyte antigens (HLAs) and cell adhesion molecules in the myocardial vasculature, as well as cellular infiltration consistent with a chronic low-grade inflammation. The aim of this study was to determine whether similar histopathologic abnormalities are present in apparently healthy relatives with LVE (left ventricular enlargement).
Experimental Steps: Specimens processed for light microscopy were analyzed by an experienced cardiac histopathologist (M. J. D.), who had no knowledge of the identity and clinical details of the subjects. For immunohistochemistry, serial 4-µm frozen cryostat sections were fixed in acetone for 3 min, incubated for 30 min with mouse monoclonal antibodies to intercellular adhesion molecule-1 (ICAM-1) (CD54, clone 6.5B5, , Oxford, UK), HLA-DR (clone B.C10, ), HLA-DQ (clone SPV-L3, ), CD3 (clone UCHT1, ) and CD68 (clone EBM11, Dakopatts, Glostrup, Denmark), washed for 15 min in phosphate-buffered saline and incubated for 25 min with affinity-purified biotinylated goat antibodies to mouse immunoglobulin G (IgG) (Dakopatts) (gamma-chain specific). After further washing, the sections were incubated with fluorescein isothiocyanate-conjugated streptavidin (Dakopatts), which labels biotin at a ratio of 3 molecules of streptavidin to 1 molecule of biotin. On sections stained for adhesion molecules and HLA markers, rabbit IgG to factor VIII (Dakopatts) (endothelial cell marker) was applied for 25 min and, after repeat washing, the sections were incubated with tetraethylrhodamine isothiocyanate-conjugated goat anti-rabbit IgG (Sigma, Cambridge, UK). On sections stained for CD3 and CD68, a propidium iodide nuclear stain was applied to facilitate cell counting.A Zeiss Axioplan (Herts, UK) photomicroscope with ultraviolet epi-illumination equipped with filters for two color immunofluorescence was used. Cell counts were performed using a grid system. The count for the entire section was obtained, and the area of the specimen was measured using an Optimax (Cambridge, UK) digitizing system to determine cell counts per unit area. In accordance with published data, an inflammatory cell count >7/mm2 was considered elevated (24). Image analysis was employed to quantify adhesion molecule and class II antigen expression. Double-exposure images obtained from a Photonic-Sciences 3 chip (Millham, UK) color-cooled camera were linked to a Kontron (Munich, Germany) 400 system for computer-assisted analysis. The following measurements were obtained: 1) total area expressing factor VIII (endothelium) as a percentage of the area of the section; and 2) total area expressing both factor VIII and HLA/adhesion molecule (i.e., endothelialexpressed antigen) from double-exposed images. From these measurements, the proportion of endothelialexpressed antigen was calculated (thus controlling for potential discrepancies in the amount of endothelium present in each section). In addition, semiquantitative assessment of staining intensity was performed in a standard fashion with visual grading: negative (N); weak (1+); moderate (2+); and high (3+). All analyses were performed by an investigator blinded to the identity of the coded sections.
Number Of Protocols: 1
- Target Reference:
- 1. Beverley, P.C.L. and Callard, R.E. (1981) Distinctive functional characteristics of human T lymphocytes defined by E rosetting or a monoclonal anti T-cell antibody. Eur. J. Immunol. 11: 329-334.
2. Zarkesh-Esfahani, H. et al. (2001) High-Dose Leptin activates human leukocytes via receptor expression on monocytes. J. Immunol. 167: 4593 - 4599.
3. Kung, P. et al. (1979) Monoclonal antibodies defining distinctive human T cell surface antigens. Science. 206: 347-9.
4. Clevers, H. et al. (1988) The transmembrane orientation of the epsilon chain of the TcR/CD3 complex. Eur. J. Immunol. 18: 705-10.
5. Clark, E.A. et al. (1983) Evolution of epitopes on human and non-human primate lymphoid cell surface antigens. Immunogenetics 18: 599-615.
6. Clevers, H. et al. (1988) The T cell receptor/CD3 complex: a dynamic protein ensemble. Annu. Rev. Immunol. 6: 629-62.
7. Meuer, S.C. et al. (1983) Evidence for the T3-associated 90 k heterodimer as the T cell antigen receptor. Nature 303: 808-10.
8. Clark, E.A. et al. (1989) Leucocyte cell surface enzymology: CD45 (LCA, T200) is a protein tyrosine phosphatase. Immunology Today 10: 225-8.
9. Campana, D. et al. (1987) The cytoplasmic expression of CD3 antigens in normal and malignant cells of the T lymphoid lineage. J. Immunol. 138: 648-55.
10. Erber, W.N. et al. (1984) Immunocytochemical detection of T cell and B cell populations in routine blood smears. Lancet 1(8385): 1042-6.
11. Erber, W.N. et al. (1986) APAAP labelling of blood and bone-marrow samples for phenotyping leukaemia. Lancet 1(8485): 761-5.
12. Denning, S.M. et al. (1987) Activation of human thymocytes via CD3 and CD2 molecules. In McMichael, A.J. et al. Leucocyte Typing III: White Cell Differentiation Antigens. Edited by Oxford University Press pp 144-7.
13. Grogan, T.M. et al. (1985) Peripheral T cell lymphoma: aggressive disease with heterogeneous immunotypes. Am. J. Clin. Pathol. 83: 271-88.
14. Maggiorella, M. et al. (1998) Detection of Infectious Simian Immunodeficiency Virus in B- and T- cell lymphomas of experimentally infected macaques. Blood. 91 (9): 3103 - 3111.