Anti-CD3 Antibody (UCHT-1)
Based on 1 Customer Validation
Anti-CD3 Antibody (UCHT-1) is an antibody targeting CD3. Anti-CD3 Antibody (UCHT-1) activates the phospholipase C and phosphatidylinositol 3-kinase (PI3K) signaling pathways, elevates intracellular calcium levels, regulates the T3 epitope, and triggers T lymphocyte proliferation. Anti-CD3 Antibody (UCHT-1) also inhibits lymphocyte proliferation, abolishes IL-2 production, and blocks the expression of IL-2 receptors. Anti-CD3 Antibody (UCHT-1) is applicable to research related to graft-versus-host disease, acute myeloid leukemia, and B-cell acute lymphoblastic leukemia.
For research use only. We do not sell to patients.
- Purity : 97.26%
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
- Protocols
- Purity & Documentation
- References
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Help & FAQs
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Biological Activity
Description
Isotype
Mouse IgG1 kappa
Recommend Isotype Controls
Species Reactivity
Human
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CD3 |
IL-2 |
PI3K |
PLC |
In Vitro
Anti-CD3 Antibody (UCHT-1) (10 μg/mL) significantly elevates the intracellular free calcium concentration in freshly isolated human primary T lymphocytes from a basal level of 32.5 nM to 112 nM[1].
Anti-CD3 Antibody (UCHT-1) (10 μg/mL; 10 min) activates phosphoinositide 3-kinase in purified human primary T lymphocytes[1].
Anti-CD3 Antibody (UCHT-1) (1 μL of 1-mg/mL solution per million WBCs; ~30 min) eliminates human T cells while preserving non-T human cell populations in unfractionated human UCB WBCs[2].
Anti-CD3 Antibody (UCHT-1) (0.01-1 μg/mL; 3-24 h incubation, 1-3 days culture) induces time- and dose-dependent modulation of T3 antigenic sites on PBMC from UCHT1-nonresponsive human donors, with complete depletion at 1 μg/mL after 6 h, and T3-reexpressing cells regain partial proliferative responsiveness to PHA in parallel with T3 reexpression[3].
Anti-CD3 Antibody (UCHT-1) (1-10000 ng/mL; 1-6 days) triggers mitogenic proliferation in PBMC from 62.5% of healthy adult donors (80% male, 45% female) via an IL 2-independent mechanism, with optimal activity at 100 ng/mL over 3 days[4].
Anti-CD3 Antibody (UCHT-1) (100 ng/mL; 3 days)-induced proliferation in PBMC from responsive healthy donors at 100 ng/mL over 3 days is not inhibited by anti-Tac antibody, confirming an IL2-independent mechanism[4].
Anti-CD3 Antibody (UCHT-1) (1 μg/mL) induces a biphasic [Ca2+]i response in human peripheral blood T lymphocytes, with an initial transient peak from intracellular Ca2+ release and a sustained plateau from extracellular Ca2+ influx; the transient peak is eliminated by intracellular BAPTA buffering[5].
Anti-CD3 Antibody (UCHT-1) (0.01-10 μg/mL; 10 min pre-incubation, 3-6 days culture) potently suppresses proliferation of PBMC and T cells from UCHT1-nonresponsive human donors induced by PHA, Concanavalin A (Con A) (HY-P2149), Candidin (HY-182559), or allogeneic non-T cells[3].
Anti-CD3 Antibody (UCHT-1) (1 μg/mL; added at culture initiation or 0-48 h post-stimulation, 3-6 days culture) maximally abrogates proliferation of PHA-stimulated PBMC and allogeneic non-T cell-stimulated T cells from UCHT1-nonresponsive human donors only when added at culture initiation, with no inhibitory effect when added 24 h (PHA) or 48 h (allogeneic non-T cells) post-stimulation[3].
Anti-CD3 Antibody (UCHT-1) (1 μg/mL) almost completely abrogates IL-2 production by PBMC stimulated with PHA and T cells stimulated with allogeneic non-T cells from UCHT1-nonresponsive human donors across all tested time points[3].
Anti-CD3 Antibody (UCHT-1) (1 μg/mL; continuous incubation, 1-3 days culture) completely blocks IL-2 receptor expression on PHA-stimulated PBMC from UCHT1-nonresponsive human donors over 3 days of culture[3].
Anti-CD3 Antibody (UCHT-1) (100 ng/mL; 1-3 days) induces IL2 receptor expression in 65-72% of PBMC from responsive healthy donors after 2-3 days of incubation at 100 ng/mL, but fails to induce meaningful IL2 receptor expression in PBMC from non-responsive donors[4].
Anti-CD3 Antibody (UCHT-1) (1 μg/mL; 15 min, 1 h) induces full IL-2 secretion, IL-2 receptor expression, and proliferation in human peripheral blood mononuclear cells with extracellular Ca2+, even when intracellular Ca2+ release-mediated transient [Ca2+]i peaks are buffered by BAPTA; extracellular Ca2+ is required for IL-2 secretion and proliferation but not IL-2 receptor expression[5].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:PBMC and T cells
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Concentration:0.1 μg/mL; 1 μg/mL; 10 μg/mL
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Incubation Time:10 min pre-incubation, 3-6 days culture
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Result:Potently suppressed proliferation of PBMC and T cells from UCHT1-nonresponsive human donors induced by PHA, Concanavalin A, Candidin, or allogeneic non-T cells
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:NSGS, NRGS, NSG, NRG immunodeficient mice (xenogeneic GVHD model; preconditioned with sublethal total body irradiation or busulfan injection)[2]
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Dosage:1 μg per 1 million white blood cells (in vitro preincubation); 1 mg/kg (i.p. single dose); 10 mg/kg (i.p. single dose)
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Administration:in vitro preincubation; ~30 minutes at 4°C prior to intravenous cell injection; i.p.; single dose within 48 hours of cell injection
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Result:Eliminated detectable human CD45+CD3+ T cells in peripheral blood 2 weeks after injection.
Prevented xenogeneic GVHD-induced mortality and weight loss.
Preserved non-T human cell engraftment, primarily myeloid markers, in peripheral blood.
Gene ID
Accession
Conjugated
Unconjugated
Reconsititution
The product can be reconstituted/diluted with sterile PBS or saline.
Format
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Product Image
Application
ELISA, FACS, Functional assay
Chemical Information
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Appearance Liquid
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Color Colorless to light yellow
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Formulation
Please refer to the lot-specific COA for specific buffer information.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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CFSE Dye Dilution Proliferation Assay
The CFSE (carboxyfluorescein diacetate succinimidyl ester) dye dilution proliferation assay is based on the covalent labeling of intracellular proteins by a cell-permeant fluorescent dye that becomes fluorescent upon intracellular ester cleavage and then is stably retained within cells. As labeled cells divide, the dye is partitioned equally between daughter cells, resulting in a stepwise halving of fluorescence intensity that can be quantified by flow cytometry to determine the number of cell divisions undergone by each cell population. This fluorescence dilution approach enables quantitative tracking of lymphocyte proliferation at the single-cell level over multiple rounds of division. CFSE-based proliferation analysis has been widely applied to measure antigen-driven lymphocyte expansion in vitro, where discrete fluorescence peaks correspond to successive cell divisions and allow reconstruction of proliferative history within heterogeneous populations.
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Calcium Spark Assay
Calcium sparks are localized, transient increases in intracellular calcium concentration ([Ca2+]i) that occur in cardiac myocytes and represent elementary events underlying excitation-contraction coupling. These events are generated by the coordinated opening of clusters of ryanodine receptors (RyRs) on the sarcoplasmic reticulum membrane, leading to a brief release of Ca2+ into the cytosol. The detection and analysis of calcium sparks provide insights into the mechanisms of calcium handling and signaling in cardiac cells. Imaging techniques using fluorescent calcium indicators such as Fluo-3 are employed to visualize these subcellular calcium transients with high spatial and temporal resolution. The protocol is based on established methodologies described in primary literature for both experimental measurement and automated analysis of calcium sparks.
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Ca2+ Staining Technique
Ca2+ staining is an experimental technique that utilizes specific fluorescent probes (such as Fluo-4 AM, Fura-2, etc.) to qualitatively or quantitatively detect dynamic changes in intracellular Ca2+ concentrations; this is achieved by monitoring the changes in fluorescent signals generated when these probes bind to free intracellular calcium ions. The underlying principle relies primarily on the presence of chelating groups within the probe's molecular structure that possess high affinity for calcium ions.
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
Purity & Documentation
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Data Sheet (267 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Inhibitory Antibodies User Guide (603 KB)
References
[1]. Turner L, et al. RANTES-activated human T lymphocytes. A role for phosphoinositide 3-kinase. Journal of immunology (Baltimore, Md. : 1950). 1995 Sep 01;155(5):2437-44. [Content Brief]
[2]. Wunderlich M, et al. OKT3 prevents xenogeneic GVHD and allows reliable xenograft initiation from unfractionated human hematopoietic tissues. Blood. 2014 Jun 12;123(24):e134-44. [Content Brief]
[3]. Van Wauwe JP, et al. Inhibition of lymphocyte proliferation by monoclonal antibody directed against the T3 antigen on human T cells. Cellular immunology. 1984 Jul;86(2):525-34. [Content Brief]
[4]. Van Wauwe JP, et al. Human T lymphocyte activation by monoclonal antibodies; OKT3, but not UCHT1, triggers mitogenesis via an interleukin 2-dependent mechanism. Journal of immunology (Baltimore, Md. : 1950). 1984 Jul;133(1):129-32. [Content Brief]
[5]. Gelfand EW, et al. Uptake of extracellular Ca2+ and not recruitment from internal stores is essential for T lymphocyte proliferation. European journal of immunology. 1988 Jun;18(6):917-22. [Content Brief]
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Anti-CD3 Antibody (UCHT-1) Related Classifications
HY-P991842 Related Classifications
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