CD3 Antibody (SP7)

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441] - FFPE human tonsil stained with CD3 antibody.

Immunohistochemistry-Frozen: CD3 Antibody (SP7) [NB600-1441] - T-cells and microglia in mouse spinal cord with acute EAE. Mouse was perfused with 4% paraformaldehyde. Spinal cord was post-fixed overnight, followed by cryoprotection with 30% sucrose for 24h. Spinal cord sections were treated with antigen-retrieval buffer (pH 6.0) at 60C for 10 min, then cooled down to room temperature (this step should not be omitted). Anti-CD3 antibody was diluted at 1:200. Incubation: room temperature overnight. IHC-Fr image submitted by a verified customer review.

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441] - Mouse lung tissue stained against C3aR normal conditions (left) and knockdown conditions (right). IHC-P image submitted by a verified customer review.

• Reactivity: Hu, Mu, Po, Ca
• Applications: Flow, ICC/IF, IHC, IHC-Fr, IHC-P, KD
• Clone: SP7
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

CD3 Antibody (SP7) Summary

• Immunogen: This CD3 antibody was developed against a synthetic peptide: KAKAKPVTRGAGA, corresponding to amino acids 156-168 of Human CD3 epsilon chain.
• Epitope: aa 156-168 of epsilon chain of human CD3 protein (intracytoplasmic).
• Localization: Type I membrane protein.
• Specificity: The CD3 antigen is present on early thymocytes and mature T cells and is generally regarded as a pan-T cell marker. This antibody will help detect CD3 expression in normal and neoplastic tissues. This antibody reacts with the intracytoplasmic portion of the CD3 antigen expressed by T cells. It stains human T cells in both the cortex and medulla of the thymus and in peripheral lymphoid tissues. This antibody is suitable for staining normal and neoplastic T cells in formalin-fixed, paraffin-embedded tissues.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: CD3E
• Purity: Tissue culture supernatant

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:25-1:50
  • Immunohistochemistry-Frozen 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:25-1:50
  • Knockdown Validated
• Application Notes: IHC-P: recommended pretreatment of citrate buffer, pH 6.0. Recommended incubation time of 30-60 min at RT. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 29037255). Use in FLOW reported in scientific literature (PMID: 31079916).
• Theoretical MW: 23.1 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Human Tonsil Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:35111697). Canine reactivity per customer review. Use in Porcine reported in scientific literature (PMID:33839961)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tissue culture supernatant
• Preservative: 0.05% Sodium Azide
• Purity: Tissue culture supernatant

Alternate Names for CD3 Antibody (SP7)

• CD_antigen: CD3e
• CD3 antigen, delta subunit
• CD3
• CD3d antigen
• CD3d antigen, delta polypeptide (TiT3 complex)
• CD3d molecule, delta (CD3-TCR complex)
• CD3-DELTA
• CD3e antigen
• CD3e antigen, epsilon polypeptide (TiT3 complex)
• CD3e molecule, epsilon (CD3-TCR complex)
• CD3e
• CD3-epsilon
• CD3g antigen
• CD3g antigen, gamma polypeptide (TiT3 complex)
• CD3g molecule, epsilon (CD3-TCR complex)
• CD3g molecule, gamma (CD3-TCR complex)
• CD3G
• CD3-GAMMA
• FLJ17620
• FLJ17664
• FLJ18683
• FLJ79544
• FLJ94613
• IMD18
• MGC138597
• T3DOKT3, delta chain
• T3E
• T-cell antigen receptor complex, epsilon subunit of T3
• T-cell receptor T3 delta chain
• T-cell surface antigen T3/Leu-4 epsilon chain
• T-cell surface glycoprotein CD3 delta chain
• T-cell surface glycoprotein CD3 epsilon chain
• TCRE

Background

CD3 (cluster of differentiation marker 3) is a multi-subunit transmembrane protein that is expressed on the surface of T-cells and forms a complex with the T-cell receptor (TCR) (1-3). CD3 consists of four distinct membrane protein isoforms: CD3-delta (delta), CD3-epsilon (epsilon), CD3-gamma, and CD3-zeta (1-3). The CD3 subunits organize into a complex containing three sets of dimers: CD3-epsilondelta, CD3-epsilongamma, CD3-zetazeta. The CD3 complex binds to the TCR heterodimer (alphabeta or gammadelta) to form the transmembrane TCR-CD3 complex (2-3). Structurally, the CD3 protein chains have an extracellular region, a transmembrane domain, and a cytoplasmic trail region (2-3). The CD3-epsilondelta and CD3-epsilongamma heterodimers also contain an extracellular immunoglobulin (Ig)-like domain, classifying them as part of the immunoglobulin superfamily (2-3). The cytoplasmic tail of each CD3 chain also contains one (delta, epsilon, gamma) or three (zeta) immunoreceptor tyrosine-based activation motifs (ITAMs), for a total of 10 ITAMs in the whole CD3 complex (2-3). Following, TCR binding to peptide major histocompatibility complex (p-MHC), the CD3 ITAMs are phosphorylated by the Src kinase Lck and are important for recruiting ZAP70 and initiating TCR signaling cascade activation (2). While similar in structure, the CD3 chains vary in length and molecular weight (4). The CD3-delta is 171 amino acids (aa) in length with a theoretical molecular weight of 18.9 kDa (4, 5). The CD3-epsilon is 204 aa long and has a theoretical molecular weight of 23 kDa (4, 5). CD3-gamma is 182 aa long with a theoretical molecular weight of 20.4 kDa (4, 5). Finally, CD3-zeta is 164 aa in length with a theoretical molecular weight of 18.6 kDa (4, 5).

CD3 proteins are expressed on the surface of thymocytes during thymocyte development, proliferation, and maturation to T-cells (4, 6, 7). During T-cell development CD4-CD8- double negative (DN) cells differentiate to CD4+CD8+ double positive (DP) cells before progressing to single positive (SP) CD4+ helper T-cells or CD8+ cytotoxic T-cells (4, 6, 7). As CD3 plays an important role in thymocyte development, it is understandable that CD3 defects and mutations in CD3 protein chains cause severe combined immunodeficiencies (SCIDs) (8). Additionally, a subset of CD3+ T-cells that co-express CD20 are described in a variety of diseases including rheumatoid arthritis, multiple sclerosis, CD20+ T-cell leukemia/lymphoma, and HIV (9). Clinical trials and animal models have shown that anti-CD3 monoclonal antibodies are a promising treatment modality for inflammatory disorders and autoimmune diseases, such as type I diabetes (10).

References

1. Chetty, R., & Gatter, K. (1994). CD3: structure, function, and role of immunostaining in clinical practice. The Journal of pathology. https://doi.org/10.1002/path.1711730404

2. Mariuzza, R. A., Agnihotri, P., & Orban, J. (2020). The structural basis of T-cell receptor (TCR) activation: An enduring enigma. The Journal of biological chemistry. https://doi.org/10.1074/jbc.REV119.009411

3. Kuhns, M. S., Davis, M. M., & Garcia, K. C. (2006). Deconstructing the form and function of the TCR/CD3 complex. Immunity. https://doi.org/10.1016/j.immuni.2006.01.006

4. Clevers, H., Alarcon, B., Wileman, T., & Terhorst, C. (1988). The T cell receptor/CD3 complex: a dynamic protein ensemble. Annual review of immunology. https://doi.org/10.1146/annurev.iy.06.040188.003213

5. Uniprot: CD3-delta (P04234), CD3-epsilon (P07766), CD3-gamma (P09693), CD3-zeta (P20963)

6. D'Acquisto, F., & Crompton, T. (2011). CD3+CD4-CD8- (double negative) T cells: saviours or villains of the immune response?. Biochemical pharmacology. https://doi.org/10.1016/j.bcp.2011.05.019

7. Dave V. P. (2009). Hierarchical role of CD3 chains in thymocyte development. Immunological reviews. https://doi.org/10.1111/j.1600-065X.2009.00835.x

8. Fischer, A., de Saint Basile, G., & Le Deist, F. (2005). CD3 deficiencies. Current opinion in allergy and clinical immunology. https://doi.org/10.1097/01.all.0000191886.12645.79

9. Chen, Q., Yuan, S., Sun, H., & Peng, L. (2019). CD3+CD20+ T cells and their roles in human diseases. Human immunology. https://doi.org/10.1016/j.humimm.2019.01.001

10. Kuhn, C., & Weiner, H. L. (2016). Therapeutic anti-CD3 monoclonal antibodies: from bench to bedside. Immunotherapy. https://doi.org/10.2217/imt-2016-0049

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD3 Antibody (NB600-1441)(25)

Publications using NB600-1441

• Pilling D, Sahlberg K, Karhadkar T et al. The sialidase NEU3 promotes pulmonary fibrosis in mice bioRxiv Feb 14 2022 (ICC/IF, Mouse)
• Glennon E, Tongogara T, Primavera V et al. Elucidating Spatially-Resolved Changes in Host Signaling During Plasmodium Liver-Stage Infection Frontiers in Cellular and Infection Microbiology Jan 17 2022 [PMID: 35111697] (ICC/IF, Mouse)
• Lu BY, Gupta R, Aguirre-Ducler A Et al. Spatially resolved analysis of the T cell immune contexture in lung cancer-associated brain metastases Journal for immunotherapy of cancer Oct 1 2021 [PMID: 34670827]
• Miao T, Wang T, Feng T Et al. Activated invariant natural killer T cells infiltrate aortic tissue as key participants in abdominal aortic aneurysm pathology Immunology Aug 11 2021 [PMID: 34379797] (ICC/IF, Human)
• Fujimoto N, He Y, D'Addio M et al. Single-cell mapping reveals new markers and functions of lymphatic endothelial cells in lymph nodes PLoS Biol. Apr 1 2020 [PMID: 32251437] (IHC, Mouse)
• Yotis DY DM Differential Effects of KIM-1 in Subcutaneous and Orthotopic Renca Models of Kidney Cancer Thesis Jan 1 2021
• Maisonneuve C, Tsang D, Foerster E Et Al. Nod1 Promotes Colorectal Carcinogenesis by Regulating the Immunosuppressive Functions of Tumor-Infiltrating Myeloid Cells SSRN Journal Jun 26 2020
• Ji Y, Fan X, Zhang Y et al. Glycine regulates mucosal immunity and the intestinal microbial composition in weaned piglets Amino acids Apr 11 2021 [PMID: 33839961] (IHC, Porcine)
• Ingelfinger F, Krishnarajah S, Kramer M et al. Single-cell profiling of myasthenia gravis identifies a pathogenic T cell signature Acta neuropathologica Mar 28 2021 [PMID: 33774709] (IHC-Fr, Human)
• Martinez-Morilla S, Zugazagoitia J, Wong PF, et al. Quantitative analysis of CMTM6 expression in tumor microenvironment in metastatic melanoma and association with outcome on immunotherapy Oncoimmunology Dec 29 2020 [PMID: 33457084] (IHC-P, Human)
• Weiss SA, Zito C, Tran T et al. Melanoma brain metastases have lower T-cell content and microvessel density compared to matched extracranial metastases J. Neurooncol. Sep 24 2020 [PMID: 32974852] (IHC, Human)
• Lee H, Lee YS, Harenda Q et al. Beta Cell Dedifferentiation Induced by IRE1 alpha Deletion Prevents Type 1 Diabetes Cell Metab. Apr 7 2020 [PMID: 32220307]
• Scortegagna M, Hockemeyer K, Dolgalev I et al. Siah2 control of T-regulatory cells limits anti-tumor immunity Nat Commun Jan 7 2020 [PMID: 31911617] (ICC/IF, Mouse)
• Manogaran P, Samardzija M, Schad AN et al. Retinal pathology in experimental optic neuritis is characterized by retrograde degeneration and gliosis Acta Neuropathol Commun Jul 17 2019 [PMID: 31315675] (IHC, Mouse)
• Toki MI, Merritt CR, Wong PF, et al. High-plex predictive marker discovery for melanoma immunotherapy treated patients using Digital Spatial Profiling Clin. Cancer Res. 2019 Jun 12 [PMID: 31189645] (ICC/IF, Human)
• Scheidecker S, Bar S, Stoetzel C et al. Fate-Mapping of GM-CSF Expression Identifies a Discrete Subset of Inflammation-Driving T Helper Cells Regulated by Cytokines IL-23 and IL-1 beta Immunity May 21 2019 [PMID: 31079916] (Flow, ICC/IF, Mouse)
• Du H, Hirabayashi K, Ahn S et al. Antitumor Responses in the Absence of Toxicity in Solid Tumors by Targeting B7-H3 via Chimeric Antigen Receptor T Cells Cancer Cell Feb 11 2019 [PMID: 30753824] (IHC, Human)
• Hartwig T, Zwicky P, Schreiner B et al. Regulatory T Cells Restrain Pathogenic T Helper Cells during Skin Inflammation Cell Rep Dec 26 2018 [PMID: 30590032] (ICC/IF, Human)
• Toki MI, Mani N, Smithy JW et al. Immune marker profiling and PD-L1 expression across Non-Small Cell Lung Cancer mutations. J Thorac Oncol. Sep 26 2018 [PMID: 30267840] (ICC/IF, Human)
• Gushchina S, Pryce G, Yip PK et al. Increased expression of colony-stimulating factor-1 in mouse spinal cord with experimental autoimmune encephalomyelitis correlates with microglial activation and neuronal loss Glia Aug 25 2018 [PMID: 30144320] (IHC-Fr, Mouse)
• Teillaud JL, Regard L, Martin C et al. Exploring the Role of Tertiary Lymphoid Structures Using a Mouse Model of Bacteria-Infected Lungs Methods Mol. Biol. Aug 25 2018 [PMID: 30141016] (IHC-P, Mouse)
• Gettinger SN, Choi J, Mani N et al. A dormant TIL phenotype defines non-small cell lung carcinomas sensitive to immune checkpoint blockers Nat Commun Aug 10 2018 [PMID: 30097571] (IHC-P, Human)
• Carvajal-Hausdorf DE, Mani N, Velcheti V et al. Objective measurement and clinical significance of IDO1 protein in hormone receptor-positive breast cancer. J Immunother Cancer. 2017 Oct 17 [PMID: 29037255] (ICC/IF, Human)
• England CG, Jiang D, Ehlerding EB et al. (89)Zr-labeled nivolumab for imaging of T-cell infiltration in a humanized murine model of lung cancer. Eur. J. Nucl. Med. Mol. Imaging 2017 Aug 19 [PMID: 28821924] (IHC, Mouse)
• Brown J, Wimberly H, Lannin DR et al. Multiplexed Quantitative Analysis of CD3, CD8, and CD20 Predicts Response to Neoadjuvant Chemotherapy in Breast Cancer. Clin Cancer Res. 2014 Sep 25 [PMID: 25255793] (IHC, Human)

Reviews for CD3 Antibody (NB600-1441) (3)

Average Rating: 5

(Based on 3 reviews)

We have 3 reviews tested in 3 species: Human, Mouse, Canine and Human.

reviewed by: Tyler Crowe IHC-Fr Human 08/20/2018

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Frozen human skin sections
• Species: Human
• Lot: J275

Comments

• Comments: Human skin section fixed with PFA, and permeablized with 0.5% Triton. Sections were blocked with 5% BSA, before anti-CD3 ab was applied at 1:100 for 1hr at 37C. anti-rb-A568 secondary was used, and counterstained with DAPI

reviewed by: Xuenong Bo IHC-Fr Mouse 02/21/2018

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: mouse spinal cord cryo-sections
• Species: Mouse
• Lot: H024

Comments

• Comments: Mouse was perfused with 4% paraformaldehyde. Spinal cord was post-fixed overnight, followed by cryoprotection with 30% sucrose for 24h. Spinal cord sections were treated with antigen-retrieval buffer (pH 6.0) at 60C for 10 min, then cooled down to room temperature (this step should not be omitted). Anti-CD3 antibody was diluted at 1:200. Incubation: room temperature overnight.

reviewed by: Hans Snyder IHC-P Canine and Human 08/25/2017

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Human Tonsil & Canine lymph paraffin slides
• Species: Canine and Human
• Lot: H024

Comments

• Comments: Works great in both Human and canine tissues.
  Sodium citrate pH 6.0
  Concentration 1:100

Bioinformatics

• Gene Symbol: CD3E
• Entrez:
  • 915(Human)
  • 916(Human)
  • 917(Human)