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Alexa Fluor® 647 anti-mouse CD3ε Antibody

Pricing & Availability
Clone
145-2C11 (See other available formats)
Regulatory Status
RUO
Other Names
CD3ε, T3, CD3
Isotype
Armenian Hamster IgG
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Product Citations
publications
145-2C11_A647_CD3e_Antibody_2_092021.png
Paraformaldehyde-fixed (4%), 500 μm-thick mouse spleen section was processed according to the Ce3DTM Tissue Clearing Kit protocol (cat. no. 427701). The section was costained with anti-mouse/human CD45R/B220 Antibody (clone RA3-6B2) Alexa Fluor® 488 at 5 µg/mL (green), anti-mouse CD68 Antibody (clone FA-11) Alexa Fluor® 594 at 5 µg/mL (blue), and anti-mouse CD3ε Antibody (clone 145-2C11) Alexa Fluor® 647 at 5 µg/mL (magenta). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 10X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
Watch the video.
  • 145-2C11_A647_CD3e_Antibody_2_092021.png
    Paraformaldehyde-fixed (4%), 500 μm-thick mouse spleen section was processed according to the Ce3DTM Tissue Clearing Kit protocol (cat. no. 427701). The section was costained with anti-mouse/human CD45R/B220 Antibody (clone RA3-6B2) Alexa Fluor® 488 at 5 µg/mL (green), anti-mouse CD68 Antibody (clone FA-11) Alexa Fluor® 594 at 5 µg/mL (blue), and anti-mouse CD3ε Antibody (clone 145-2C11) Alexa Fluor® 647 at 5 µg/mL (magenta). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 10X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
    Watch the video.
  • 145-2C11_Alx647_030206
    C57BL/6 mouse splenocytes were stained with CD3e (clone 145-2C11) Alexa Fluor® 647 (filled histogram) or Armenian hamster IgG Alexa Fluor® 647 isotype control (open histogram).
  • 145-2C11_Alx647_2_030206
    C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then the section was stained with 10 µg/ml of CD3ε (clone 145-2c11) Alexa Fluor® 647 (red), and B220 (clone RA3-6B2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured by 10X objective.
Compare all formats See Alexa Fluor® 647 spectral data
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100324 25 µg £70
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100322 100 µg £157
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Description

CD3ε is a 20 kD transmembrane protein, also known as CD3 or T3. It is a member of the Ig superfamily and primarily expressed on T cells, NK-T cells, and at different levels on thymocytes during T cell differentiation. CD3ε forms a TCR complex by associating with the CD3δ, γ and ζ chains, as well as the TCR α/β or γ/δ chains. CD3 plays a critical role in TCR signal transduction, T cell activation, and antigen recognition by binding the peptide/MHC antigen complex.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Armenian Hamster
Immunogen
H-2Kb-specific mouse cytotoxic T lymphocyte clone BM10-37
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
IHC-F, 3D IHC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µl volume. For immunohistochemistry on frozen tissue sections, a concentration range of 5.0 - 10.0 µg/ml is suggested. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

Clone 145-2C11 is useful for in vitro blocking of target-specific CTL-mediated cell lysis1, as well as T cell activation assays, inducing proliferation and cytokine production1,2,7,12,16. It also induces apoptosis in immature thymocytes32,  and in vivo T cell depletion8-10. Additional reported applications (for relevant formats of this clone) include: immunoprecipitation1, immunohistochemical staining14,15 of acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections, Western blotting4, complement-mediated cytotoxicity6, in vitro and in vivo stimulation of T cells1,2,7,12,16, immunofluorescent staining5, and in vivo T cell depletion8-10. The 145-2C11 antibody has been reported to block the binding of 17A2 antibody to CD3 epsilon-specific T cells11. Clone 145-2C11 is not recommended for formalin-fixed paraffin embedded sections. The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 100314). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 100340) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg).

Application References

(PubMed link indicates BioLegend citation)
  1. Leo O, et al. 1987. P. Natl. Acad. Sci. USA 84:1374. (IP, Activ, Block)
  2. Kruisbeek AM, et al. 1991. In Current Protocols in Immunology. 3.12.1. (Activ)
  3. Duke RC, et al. 1995. Current Protocols in Immunology. 3.17.1.
  4. Salvadori S, et al. 1994. J. Immunol. 153:5176. (WB)
  5. Payer E, et al. 1991. J. Immunol. 146:2536. (IF)
  6. Jacobs H, et al. 1994. Eur. J. Immunol. 24:934. (CMCD)
  7. Vossen ACTM, et al. 1995. Eur. J. Immunol. 25:1492. (Activ)
  8. Henrickson M, et al. 1995. Transplantation 60:828. (Deplete)
  9. Kinnaert P, et al. 1996. Transpl. Int. 9:386. (Deplete)
  10. Han WR, et al. 1999. Transpl. Immunol. 7:207. (Deplete)
  11. Miescher GC, et al. 1989. Immunol. Lett. 23:113. (Block)
  12. Terrazas LI, et al. 2005. Intl. J. Parasitology. 35:1349. (Activ)
  13. Ko SY, et al. 2005. J. Immunol. 175:3309.
  14. Podd BS, et al. 2006. J. Immunol. 176:6532. (IHC-F)
  15. Tilley SL, et al. 2007. J. Immunol. 178:3208. (IHC-F)
  16. Wang W, et al. 2007. J. Immunol. 178:4885. (Activ)
  17. Xiao S, et al. 2007. J. Exp. Med. 204:1691.
  18. Chappaz S, et al. 2007. Blood doi:10.1182/blood-2007-02-074245. (FC) PubMed.
  19. Curtsinger JM, et al.2005. J. Immunol. 175:4392. PubMed
  20. Guo Y, et al. 2008. Blood 112:480. PubMed
  21. Kenna TJ, et al. 2008. Blood 111:2091.
  22. Perchonock CE, et al. 2007. J. Immunol. 179:1768. PubMed
  23. Perchonock GE, et al. 2006. Mol. Cell. Biol. 26:6005. PubMed
  24. Kanaya T, et al. 2008. Am. J. Physiol. Gastrointest. Liver Physiol. 295:G273. PubMed
  25. de Koning BA, et al. 2006. Int. Immunol. 18:941. PubMed
  26. Schulteis RD, et al. 2008. Blood 295:G273. PubMed
  27. Qi Q, et al. 2009. Blood 114:564. PubMed
  28. Helmersson S, et al. 2013. Am J Pathol. 9440:123. Pubmed
  29. Wu S, et al. 2014. Clin Vaccine Immunol. 21:156. PubMed
  30. Yan J, et al. 2014. Vaccine. 32:2833. PubMed
  31. Guiterrez DA, et al. 2014. Diaebetes. 63:3827. PubMed
  32. Shi YF, et al. 1991. J Immunol. 146:3340. (Apop)
Product Citations
  1. Shenoy AT, et al. 2022. Cytometry A. 101:892. PubMed
  2. Yeh CH, et al. 2022. Immunity. 55:272. PubMed
  3. Tran NT, et al. 2019. Cell Rep. 28:3510. PubMed
  4. Periasamy S, et al. 2017. Nat Commun. 8:15564. PubMed
  5. Rattan A, et al. 2017. PLoS Pathog. 13:e1006248. PubMed
  6. She L, et al. 2021. JCI Insight. 6:e143509. PubMed
  7. Ly A, et al. 2020. Cell Reports. 29(8):2257-2269.e6.. PubMed
  8. Zhong W, et al. 2022. Nat Commun. 13:4390. PubMed
  9. Werbner M, et al. 2019. mSystems. 4:e00292-18. PubMed
  10. Titelbaum M, et al. 2021. iScience. 24:103093. PubMed
  11. She L, et al. 2020. PLoS One. 15:e0236744. PubMed
  12. Balzano M et al. 2019. Cell reports. 26(12):3257-3271 . PubMed
  13. Dholakia J, et al. 2022. Gynecol Oncol. 164:170. PubMed
  14. Isvoranu G, et al. 2019. Oncol Lett. 17:4197. PubMed
  15. Hendrikx S et al. 2019. Cell reports. 26(5):1227-1241 . PubMed
  16. Terlizzi M, et al. 2021. Cell Physiol Biochem. 55:539. PubMed
  17. Tsai S, et al. 2018. Cell Metab. 28:922. PubMed
RRID
AB_492861 (BioLegend Cat. No. 100324)
AB_492861 (BioLegend Cat. No. 100322)

Antigen Details

Structure
Ig superfamily, forms CD3/TCR complex with CD3δ, γ and ζ subunits and TCR (α/β and γ/δ), 20 kD
Distribution

Thymocytes (differentiation dependent), mature T cells, NK-T cells

Function
TCR signal transduction, T cell activation, antigen recognition
Ligand/Receptor
Peptide antigen/MHC-complex
Cell Type
NKT cells, T cells, Thymocytes, Tregs
Biology Area
Immunology
Molecular Family
CD Molecules, TCRs
Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Davis MM. 1990. Annu. Rev. Biochem. 59:475.
3. Weiss A, et al. 1994. Cell 76:263.

Gene ID
12501 View all products for this Gene ID
UniProt
View information about CD3epsilon on UniProt.org
Go To Top Version: 2    Revision Date: 09/20/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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