Key Spec Table
|Species Reactivity||Key Applications||Host||Format||Antibody Type|
|H||FC, IHC||M||FITC||Monoclonal Antibody|
|Description||Anti-CD3 Antibody, clone UCHT1, FITC conjugated|
|Presentation||The conjugate is supplied as a 100 test/1 mL vial in phosphate buffered saline containing 10mM sodium azide and 1mg/ml bovine serum albumin. For flow cytometry we recommend using 10μL of conjugate per test.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Store at +4°C protected from light. DO NOT FREEZE. For long term use and storage aliquot conjugate into small volumes and store at +4°C.|
|Material Size||100 assays|
|Anti-CD3, clone UCHT1, FITC conjugated - CBL150F||CBL150F|
|Anti-CD3, clone UCHT1, FITC conjugated - 2367811||2367811|
|Reference overview||Pub Med ID|
|Leukocyte cell surface enzymology: CD45 (LCA, T200) is a protein tyrosine phosphatase.|
Clark, E A and Ledbetter, J A
Immunol. Today, 10: 225-8 (1989) 1989
During 1987, striking advances were made in defining the receptors and ligands for cell-to-cell adhesion interactions involving leukocytes. In 1988, two major leukocyte differentiation antigens, CD10 (cALLA) and CD45 (LCA, T200), were shown to be enzymes while two other markers, CD4 and CD8, were found to be associated with an enzyme. In this article, Ed Clark and Jeff Ledbetter discuss recent findings in the emerging area of leukocyte cell surface enzymology with emphasis on CD45, a membrane-associated protein tyrosine phosphatase (PTPase)2,3.
|The transmembrane orientation of the epsilon chain of the TcR/CD3 complex.|
Clevers, H, et al.
Eur. J. Immunol., 18: 705-10 (1988) 1988
The antigen receptor of the T lymphocytes is one of the most complex eukaryotic membrane structures studied to date. The T cell receptor (TcR) consists of two disulfide-linked glycoprotein chains (alpha/beta or gamma/delta) and is noncovalently associated with a group of small and invariable CD 3 proteins. Four CD 3 chains have been recognized: two highly homologous glycoproteins CD 3 gamma and delta, the more distantly related nonglycosylated CD 3 epsilon chain, and the nonglycosylated CD 3 zeta, the latter being present as a homodimer. The unraveling of the architecture of the TcR/CD 3 complex is crucial to our understanding of the processes underlying its assembly, recognition and transmembrane signaling. The transmembrane orientation of the TcR chains and of CD 3 gamma and CD 3 delta can be directly inferred from their primary structure, based on the presence of concensus N-linked glycosylation sites N-terminal of their transmembrane domains. This prediction can not be made, however, for nonglycosylated molecules like the CD 3 epsilon chain. In order to determine the transmembrane orientation of CD 3 epsilon, anti-peptide antisera directed against the N-termini of the human and murine CD 3 epsilon chains were generated in rabbits. Both antisera stained intact T cells, demonstrating that the N-terminus of the CD 3 epsilon chain was located at the outer surface of the plasma membrane. The anti-human CD 3 epsilon peptide antiserum was found to be mitogenic for peripheral blood T cells, a finding previously reported only for monoclonal anti-TcR/CD 3 reagents. Using a novel transient expression system in murine T lymphocytes, the human CD 3 epsilon chain could be expressed on the surface of CD 3+, but not CD 3- murine T cells, as indicated by fluorescence staining with the anti-peptide antiserum. This experiment confirmed the specificity of the anti-peptide antiserum and, perhaps more importantly, indicated that the human CD 3 epsilon chain was correctly assembled in the murine CD 3 complex. Moreover, the anti-human CD 3 monoclonal antibody UCHT1 was found to stain T cells expressing the human CD 3 epsilon chain.
|The T cell receptor/CD3 complex: a dynamic protein ensemble.|
Clevers, H, et al.
Annu. Rev. Immunol., 6: 629-62 (1988) 1988
|Intrathymic ontogeny of the T cell receptor associated CD3 (T3) antigen.|
Swerdlow, S H, et al.
Lab. Invest., 58: 421-7 (1988) 1988
Intrathymic T cell maturation has been studied in great detail, yet much of this process remains controversial. One of the critical steps in thymocyte maturation is the surface expression of the T cell antigen receptor associated CD3 (T3) antigen. The expression and ultrastructural localization of this antigen was therefore studied in tissue sections of five human thymuses using the monoclonal antibody UCHT-1. Thymocytes were also independently categorized morphologically as small nonblastic, large blastic, or intermediate. CD3 positivity was identified in 89% of cortical thymocytes and 92% of medullary thymocytes. Of the CD3+ cells, staining was present in the perinuclear region in 48% of cortical thymocytes and in only 11% of medullary thymocytes. Surface membrane staining was present in 87% of CD3+ cortical thymocytes and 99% of CD3+ medullary thymocytes. Of the CD3+ cortical large blastic thymocytes, 97% had perinuclear staining, but only 31% had surface positivity. Conversely, 98% of CD3+ medullary large blastic thymocytes had surface staining, but only 17% had perinuclear positivity. Almost all cortical and medullary small nonblastic thymocytes had surface membrane CD3 positivity, whereas perinuclear positivity was rare in these cells. These data suggest that the most immature CD3+ thymocytes (the large blasts with perinuclear CD3 positivity only) are present almost exclusively in the cortex. Maturation defined by the loss of perinuclear CD3 positivity and the acquisition of surface CD3 also occurs in the cortex. The vast majority of medullary thymocytes have surface CD3 positivity only.
|The cytoplasmic expression of CD3 antigens in normal and malignant cells of the T lymphoid lineage.|
Campana, D, et al.
J. Immunol., 138: 648-55 (1987) 1987
Anti-CD3 (T3) Ab reacting with different proportions of thymocytes (anti-CD3a: UCHT1, anti-CD3b: T10B9, and anti-CD3c: OKT3) were tested for cytoplasmic (cCD3) and membrane (mCD3) expression in the bone marrow, thymus, and blood in man and selected primates. The expression of cCD3a and cCD3c in the perinuclear and Golgi area of large, BrdU-incorporating, strongly TdT+ thymic blasts probably represents one of the earliest signs of T cell commitment, because these blast cells are CD1-, CD4-, CD8-, and mCD3-. The cCD3+, TdT+ cells are normally restricted to the thymus and are absent among the TdT+ cells of bone marrow. The anti-CD3b Ab used, T10B9, co-caps and co-modulates with the other anti-CD3 Ab and is a T cell-specific reagent at a membrane level but does not bind to perinuclear cCD3. Instead, this reagent cross-reacts with a filamentous cytoplasmic network in non-T cells in man and in primates S. oedipus and M. rhesus despite their T cell negativity for mCD3. The characteristics of all T-ALL cases studied: cCD3+, CD7+ along with nuclear TdT+ suggest lineage fidelity to early thymic blasts. As a marked contrast, cCD3 is absent in common ALL and in AML, including cases that concomitantly express CD7 and myeloid antigens. Thus, the cCD3, TdT combination provides a very sensitive assay for residual T-ALL blasts outside the normal thymus.
|The immunocytochemical distribution of leukocytic subpopulations in human endometrium.|
Kamat, B R and Isaacson, P G
Am. J. Pathol., 127: 66-73 (1987) 1987
Thirty human endometria were selected from women aged 21-54 years who had undergone routine dilation and curettage procedures for tubal ligation, infertility dating, and irregular menstrual cycling. Histologic sections of the cases chosen were examined to exclude any major pathologic condition (including chronic endometritis). The specimens were stained with monoclonal antibodies to a common leukocytic antigen (H Leu-1 and PD7/26), pan-T-cell antigen (UCHT1), T helper/inducer and T suppressor/cytotoxic antigens (Leu-3a and UCHT4, respectively), pan-B cell antigen (To15 and Leu-12), and macrophage antigens (UCHM1 and Leu-M3). Other antibodies used included TAL-1B5 (anti-HLA-DR), Leu-7 (natural killer cell) and Na 1/34 (anti-T6/Langerhans/interdigitating reticulum cell). The endometria contained significant numbers of common leukocyte antigen-positive cells (occupying approximately 10-15% of the stroma), the numbers of which appeared to increase in the late secretory/pre-menstrual phase (20-25% of the stroma). The major leukocyte populations were T cells and macrophages; the latter, with neutrophils, appeared to account for the premenstrual increase in leukocytes. T cells were distributed both diffusely in the stroma and in periglandular stromal aggregates closely applied to the glands. The T8+ suppressor/cytotoxic population was predominant within the stromal nodules. In addition, scattered intraepithelial T suppressor/cytotoxic cells were present. Macrophages (UCHM1 and HLA-DR+) were also distributed diffusely in the stroma and as part of the periglandular stromal aggregates, in areas sending long cell processes into the epithelium. B cells appeared to be limited to scattered cells in the stroma, only increasing in number within lymphoid follicles. Natural killer cells, as defined by Leu-7+ cells, were also present, scattered singly in the stroma and within lymphoid follicles. The demonstration of large mononuclear dendritic-appearing Na 1/34+ cells within the glands of the endometrium in 5/30 cases suggests the presence of T6+ Langerhans/interdigitating reticulum cells in the endometrium. Thus, the normal endometrium has an important population of immunologically competent cells. Further study of these cell populations may elucidate their contribution, if any, to pathologic conditions in the endometrium.
|APAAP labelling of blood and bone-marrow samples for phenotyping leukaemia.|
Erber, W N, et al.
Lancet, 1: 761-5 (1986) 1986
The reliability of labelling routinely prepared air-dried peripheral-blood and bone-marrow smears with the alkaline-phosphatase/anti-alkaline-phosphatase (APAAP) method for phenotyping leukaemias was evaluated in 259 cases of haematological malignancy. All the cellular markers currently used in leukaemia diagnosis could be detected with this technique. Since the technique can be used on blood and marrow smears even after storage or postal transport, it is suggested that APAAP labelling should become the method of choice for immunophenotyping neoplastic samples in many haematology laboratories not equipped with specialised immunological diagnostic facilities. The method should also be valuable for epidemiological studies of leukaemia.
|Evidence for the T3-associated 90K heterodimer as the T-cell antigen receptor.|
Meuer, S C, et al.
Nature, 303: 808-10 (1983) 1983
Several surface molecules appear to be involved in antigen recognition by human T lymphocytes including the monomorphic 20/25K T3 structure present on all mature T lymphocytes and the subset-specific associative recognition elements, T4 and T8 (refs 1-8). More recently, Ti1, a clonally unique antigen recognition structure comprised of a 49,000 molecular weight (49K) alpha-chain and a 43K beta-chain, linked to T3 was identified on a major histocompatibility complex (MHC) class I specific T8+ T-cell clone, CT8III (ref. 9). To determine whether analogous receptor molecules could be found on other T-cell clones of differing specificity, we produced monoclonal antibodies against a clonal structure (Ti2) on an MHC class II specific T4+ lymphocyte, CT4II, derived from the same donor as CT8III. The Ti2 structure on CT4II is shown here to be a disulphide-linked heterodimer like Ti1 on CT8III and is composed of subunits of similar molecular weight. Monoclonal antibodies against Ti2 or Ti1 block antigen specific functions of the respective clone without showing any cross-reactivity. These findings suggest that each T lymphocyte, regardless of subset derivation or specificity, uses an analogous Ti heterodimer for antigen specific function. The latter is linked to T3 and expressed on the cell surface at an identical density (30,000-40,000 sites per cell).
|Distinctive functional characteristics of human "T" lymphocytes defined by E rosetting or a monoclonal anti-T cell antibody.|
Beverley, P C and Callard, R E
Eur. J. Immunol., 11: 329-34 (1981) 1981
|Monoclonal antibodies defining distinctive human T cell surface antigens.|
Kung, P, et al.
Science, 206: 347-9 (1979) 1979
Three novel nonoclonal antibodies (designed OKT1, OKT3, and OKT4) were generated against surface determinants of human peripheral T cells. Both OKT1 and OKT3 reacted with all human peripheral T cells and 5 to 10 percent of thymocytes but differed in their reactivities with T cel- lines. By contrast, OKT4 reacted with 55 percent of human peripheral T cells and 80 percent of thymocyted in that they did not react with normal B cells, null cells, monocytes, or granulocytes.
|Anti-CD3, Clone UCHT1, FITC Conjugated - Data Sheet|