Anti-Mouse CD3e Monoclonal Antibodies for Flow Cytometry

PA007524.r2a: Recombinant Anti-mouse CD3e Monoclonal Antibody (Clone: KT3), Rat IgG2a Kappa

Recombinant Rat IgG2a Monoclonal Antibody.
Clone: KT3.
Isotype: Rat IgG2a Kappa.
Source: The anti-mouse CD3e monoclonal antibody (clone: KT3) was produced in mammalian cells.
Conjugation: non-conjugated.
Specificity/Sensitivity: The recombinant rat monoclonal antibody (clone: KT3) specifically binds to the mouse CD3e.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F).
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions and HPLC.

PA007524.h1Fs: Recombinant Anti-mouse CD3e Monoclonal Antibody (Clone: KT3), Human IgG1 Fc Silent Kappa

Recombinant Human IgG1 Monoclonal Antibody.
Clone: KT3.
Isotype: Human IgG1 Fc Silent Kappa.
Source: The anti-mouse CD3e monoclonal antibody (clone: KT3) was produced in mammalian cells.
Conjugation: non-conjugated.
Specificity/Sensitivity: The recombinant rat monoclonal antibody (clone: KT3) specifically binds to the mouse CD3e.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F).
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions and HPLC.

Shipping: The antibody is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, 2 to 8°C as supplied. Do not freeze it.

References of anti-mouse CD3e antibody (KT3)

CD1-restricted CD4+ T cells in major histocompatibility complex class II-deficient mice
Cardell S, et al. J Exp Med. 1995 Oct 1;182(4):993-1004. doi: 10.1084/jem.182.4.993. PMID: 7561702
Single cell suspensions were stained with mAbs of the following specificities: KT3 (anti-mouse CD3e antibody), KT4 (V134), MR9-4 (V135), 44.22.1 (V136), KJ16 (V138.1/8.2), RR3.15 (V1311), IM7 (CD44), H57-597 (TCR??), for references see (1), MR10-2 (V139 [8]), B21.5 (V110 [9]), B20.1.1 (V?12 [10]), KT50 (V?8 [11]), M17/4 (LFA-1? [12]), YN1/1.7.4 (ICAM-1/CD54), MEL-14 (L-selectin/CD62L [13]), 16A (CD45RB [14]), PC61 (CD25 [15]), PK136 (NK1.1 [16]), and 12-15A-biotin (CD2 [17]). The sorted CD4+ cells were stimulated at 1.5-3 X 104 cells per well in anti-CD3-coated (KT3 antibody coated at 5 ?g/ml or as indicated) round-bottom, 96-well plates in Hepes-buffered RPMI 1640 medium containing glutamine supplemented with 10% FCS, 20 ?M 2-ME, 1 mM Na-pyruvate, and penicillin/streptomycin. Labeled cells were analyzed on FACScan?, ODAM ATC 3000 (Bruker Spectrospin, Wissembourg, Germany), or Coulter Epic Elite (Coulter Corp., Hialeah, FL) flow cytometers, and sorted using the ATC or Elite. Lymph node cells from wild-type, class II-deficient (II?) and ?2m/class II double-deficient (II? ?2m?) littermate mice were stained for the CD4 and CD8 markers. On the indicated days, spleen cells were prepared from mixed litters of II? and wild-type heterozygote control mice, and they were analyzed for CD4, CD8, and CD3 expression.
Tags: anti-mouse CD3e KT3; anti-mouse CD3e KT3 for flow cytometry

Anti-CD3 therapy permits regulatory T cells to surmount T cell receptor-specified peripheral niche constraints
Chatenoud L, et al. J Exp Med. 2010 Aug 2;207(8):1593-8. doi: 10.1084/jem.20100798. PMID: 20679403
For anti-CD3 treatment, mice were injected i.v. with 3 ?g/g BW of anti-CD3 mAb (clone; KT3) for 5 consecutive days. (E) Splenic CD4 + cells from BR mice 6 d after treatment with anti-CD3 or Rat IgG were analyzed by flow cytometry. (A) CD4 + cells in the spleen, pancreatic, inguinal, and mesenteric lymph node (SPL, PLN, ILN, and MLN, respectively) from mice untreated or treated with anti-CD3 or control IgG were analyzed by flow cytometry. (B) BrdU incorporation by T reg and T conv cell CD4 + splenocytes in BR mice during a 6-h pulse, before or at 3, 6, or 9 d of anti-CD3 treatment. Cells from the SPL, axillary lymph node (AxLN), MLN, and PLN of NOD.RAG o/o hosts 14 d after transfer of GFP ? cells from BDC2.5.CD90.1.Foxp3 GFP mice were analyzed by flow cytometry.
Tags: function of anti-mouse CD3e KT3; bioactivity of anti-mouse CD3e KT3

Visualizing Spatial and Stoichiometric Barriers to Bispecific T-cell Engager Efficacy
You R, et al. Cancer Immunol Res. 2022 Jun 3;10(6):702-714. doi: 10.1158/2326-6066.CIR-21-0594. PMID: 35413104
The mouse surrogate CD3 engager moiety, KE3, was based on a rat anti-mouse CD3? scFv derived from the KT3 monoclonal antibody (anti-mouse CD3e) sequence. Flow cytometry analysis of the coupling between mouse CD8 + T cells (violet proliferation dye (VPD)-labeled) and PyMT-Ctrl (control) or PyMT-EGFRvIII-Hi cells (CMTMR-labeled) co-cultured for 1 hour in the presence of BiTEs as indicated. Flow cytometry analysis of mouse CD8 + T-cell proliferation (indicated by VPD dilution), activation (indicated by surface CD25 and CD69 expression), and tumor killing induced by BiTEs when T cells were co-cultured with PyMT-EGFRvIII-Hi cells for 48 hours in the presence of different concentrations of BiTEs as indicated. Flow cytometry analysis of CD8 + T cells in the indicated groups. T cells were harvested on day 19 post tumor injection. Representative flow cytometry plots and quantification of activation markers expressed by CD8 + T cells in the indicated tumor groups.
Tags: anti-mouse CD3e KT3 antibody of low endotoxin; anti-mouse CD3e antibody (clone KT3)

Site-directed multivalent conjugation of antibodies to ubiquitinated payloads
el Hebieshy AF, et al. Nat Biomed Eng. 2025 Feb;9(2):234-248. doi: 10.1038/s41551-024-01342-z. PMID: 40204992
For the initial conjugation reaction, the KT3 hybridoma-derived anti-mouse CD3 Fab-Ub(K48R) don, the chemically synthesized Ub acc-?GG carrying an N-terminal rhodamine fluorophore (Rho-Ub acc-?GG), in combination with recombinant E1 and the lysine-48 (K48)-specific ubiquitin E2?E3 fusion protein gp78RING-Ube2g2 (ref. 26) were chosen (Fig. 1b). The following hybridoma cell lines were modified for the stable expression of ubi-tagged antibodies or antibody fragments: KT3 (kindly provided by Dr Ramon Arens (LUMC, the Netherlands)), NLDC-145 (ATCC HB-290) and TA99 (ATCC HB-8704). Next, we used flow cytometry to compare the staining of CD3 + mouse splenocytes with anti-mCD3 Rho-Ub 2-Fab to staining with fluorescein isothiocyanate (FITC)-labelled parental antibody (Fig. 1e). Flow cytometry analysis revealed a ubi-tagged TRP1/mCD3 BiTE dose-dependent T-cell activation as measured by the increased expression of Ki67, Granzyme B, CD69 and 4-1BB (CD137) by the T cells, but only when co-cultured with KPC3-TRP1 cells and not with KPC3 WT cells (Fig. 3d?g and Supplementary Fig. 8a). Division index obtained by flow cytometry analysis ( n = 4) of OT-I cells isolated from spleen. Data are shown as mean ± s.d. Unpaired t-tests, P values are noted in the figure.
Tags: anti-mouse CD3e antibody KT3; anti-CD3e (clone KT3)

Anti?CD3 treatment up?regulates programmed cell death protein?1 expression on activated effector T cells and severely impairs their inflammatory capacity
Wallberg M, et al. Immunology. 2017 Jul;151(3):260-272. doi: 10.1111/imm.12729. PMID: 28211040
The non?Fc receptor?binding anti?mouse CD3 antibody (KT3, agly?anti?CD3) was generated through genetic engineering in the Waldmann Laboratory, University of Oxford. It consists of the antigen?binding variable domain of the anti?mouse CD3 KT3 clone fused to mutated (non?FcR binding) human IgG1 heavy and ? light chains. Chimeric KT3?1.1 aglycosyl IgG1 antibody mRNA was prepared from the cells of the hybridoma KT3?1.1 and cDNA was prepared by anchor?tailed PCR. Data were collected on a Cyan Cell Cytometer (DAKO, Santa Clara, CA) and analysed using flowjo (Tree Star Inc., Ashland, OR). For intracellular cytokine staining, the cells were stimulated with PMA (50 ng/ml) and ionomycin (2000 ng/ml) for 5 hr.
Tags: clone KT3 of anti-mouse CD3e; KT3 monoclonal antibody

The Single Positive T Cells Found in CD3-?/?? /? Mice Overtly React with Self?Major Histocompatibility Complex Molecules upon Restoration of Normal Surface Density of T Cell Receptor?CD3 Complex
Lin SY, et al. J Exp Med. 1997 Feb 17;185(4):707-714. doi: 10.1084/jem.185.4.707. PMID: 9034149
For flow cytometric analyses, T cell hybridomas were first incubated with saturating concentrations of mAb directed against CD3-? (KT3), TCR-? (H57-597), TCR-? (GL3), CD4 (RL172.4), CD8 (3.168.81), or the products of various V? and V? gene segments, washed, and then stained with saturating concentration of an F(ab?)2 fragment of a FITCconjugated rabbit anti-mouse Ig (Cappel Labs., Malvern, PA). When analyzed by flow cytometry for the expression of CD4 and CD8 molecules, thymuses from 1- to 3-mo-old CD3-?/? ?/? mice were found to contain dramatically fewer single positive cells than age-matched wild-type thymuses (Fig. 1). Similar to CD3-?/? ?/? thymocytes, the single positive cells found in CD3-?/? ?/? lymph nodes expressed levels of TCR/CD3 complexes barely detectable by flow cytometry (Fig. 2; 8?10, 17). Most of the CD3 ? peripheral single positive T cells found in CD3-?/??deficient mice display constitutively upregulated levels of CD44. Lymph node cells from 3-mo-old CD3-?/? ?/? and -?/? +/+ mice were analyzed by three-color flow cytometry for CD4 and CD8 expression in combination with various cell surface markers. Fusion of CD3-?/? ?/? peripheral single positive T cells with BW ??, a CD3-? + derivative of the BW ? thymoma, results in hybridomas expressing TCR-?/? complexes at their surface. BW ?? was fused to single positive T cells isolated from the spleen and lymph nodes of 3-mo-old CD3-?/? ?/? or -?/? +/+ mice.
Tags: purified anti-mouse CD3e (KT3); anti-mouse CD3e KT3 antibody

For more references about anti-mouse CD3e antibody (KT3), please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following in vivo grade recombinant anti-human CD3 monoclonal antibodies:
Muromonab biosimilar, research grade, anti-human CD3 monoclonal antibody (Clone: OKT3)
Teplizumab biosimilar, research grade, anti-human CD3 monoclonal antibody (Clone: OKT3)
Foralumab biosimilar, research grade, anti-human CD3 monoclonal antibody (Clone: OKT3)
Recombinant Anti-human CD3 monoclonal antibody (Clone: OKT3)
Recombinant Anti-human CD3 monoclonal antibody (Clone: SP34-2)
Recombinant Anti-human CD3 monoclonal antibody (Clone: UCHT1)
Recombinant Anti-human CD3 monoclonal antibody (Clone: 12F6)

Syd Labs provides the following in vivo grade recombinant anti-human CD3 bispecific antibodies:
Recombinant Anti-human CD3 / CD3 Bispecific Antibody (Clone: OKT3 / UCHT1)
Recombinant Anti-human CD3 / CD3 Bispecific Antibody (Clone: OKT3 / SP34-2)
Recombinant Anti-human CD3 / CD3 Bispecific Antibody (Clone: SP34-2 / OKT3)

Syd Labs provides the following in vivo grade recombinant anti-mouse CD3 monoclonal antibodies:
Recombinant Anti-mouse CD3e monoclonal antibody (Clone: 145-2C11)
Recombinant Anti-mouse CD3e monoclonal antibody (Clone: 500A2)
Recombinant Anti-mouse CD3 monoclonal antibody (Clone: 17A2)
Recombinant Anti-mouse CD3e monoclonal antibody (Clone: KT3)

Syd Labs provides the following recombinant anti-human CD3 monoclonal antibodies for flow cytometry:
Recombinant Anti-human CD3 monoclonal antibody (Clone: OKT3) for flow cytometry
Recombinant Anti-human CD3 monoclonal antibody (Clone: UCHT1) for flow cytometry
Recombinant Anti-human CD3 monoclonal antibody (Clone: 12F6) for flow cytometry
Recombinant Anti-human CD3 monoclonal antibody (Clone: SP34-2) for flow cytometry

Syd Labs provides the following recombinant anti-mouse CD3 monoclonal antibodies for flow cytometry:
Recombinant Anti-mouse CD3 monoclonal antibody (Clone: 17A2) for flow cytometry
Recombinant Anti-mouse CD3e monoclonal antibody (Clone: 145-2C11) for flow cytometry
Recombinant Anti-mouse CD3e monoclonal antibody (Clone: 500A2) for flow cytometry
Recombinant Anti-mouse CD3e monoclonal antibody (Clone: KT3) for flow cytometry