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Alexa Fluor® 647 anti-mouse CD3 Antibody

Pricing & Availability
Clone
17A2 (See other available formats)
Regulatory Status
RUO
Other Names
T cell antigen receptor complex, T3
Isotype
Rat IgG2b, κ
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Product Citations
publications
1-17A2_Alx647_021606
C57BL/6 mouse splenocytes stained with 17A2 Alexa Fluor® 647
  • 1-17A2_Alx647_021606
    C57BL/6 mouse splenocytes stained with 17A2 Alexa Fluor® 647
  • 2-17A2_A647_CD3_Antibody_2_121018
    Dissected C57/B6 mouse spleen was immersed in 4% paraformaldehyde (PFA) overnight followed by 30% sucrose immersion overnight and frozen in OCT. Frozen section was blocked with 5% FBS and 5% mouse serum for 30 minutes at room temperature. Then the tissue section was stained with 5 µg/mL of anti-mouse CD3 (clone 17A2) Alexa Fluor® 647 (red) and 5 µg/mL of anti-mouse B220 (clone RA3-6B2) Alexa Fluor® 594 (blue) and 5 µg/mL of anti-mouse CD169 (clone 3D6-112) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured by 10X objective.
  • 3-17A2_A647_CD3_Antibody_3_050219
    Formalin-fixed, 400 micron-thick mouse spleen section was blocked, permeabilized and stained overnight with CD21/35 (CR2/CR1)(clone 7E9) Alexa Fluor® 594 (red), CD169 (Siglec-1) (clone 3D6.112) Alexa Fluor® 488 (green), and CD3 (clone 17A2) Alexa Fluor® 647 (blue) all at 5 µg/mL, optically cleared, then analyzed at 235 µm imaging depth on a confocal microscope. Scale bar: 100 µm. Watch the video.
  • 4-17A2_A647_CD3_Antibody_4_3D-IHC_092121.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 CD37 Antibody (clone Duno85) Alexa Fluor® 594 at 5 µg/mL (yellow), and anti-mouse CD3 Antibody (clone 17A2) 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.
Compare all formats See Alexa Fluor® 647 spectral data
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Description

CD3, also known as T3, 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. CD3 is composed of CD3ε, δ, γ and ζ chains. It forms a TCR complex by associating with 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
Rat
Immunogen
γδTCR-positive T-T hybridoma D1
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
SB - Community 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 2.5 - 5.0 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 633 nm / 635 nm.


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

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Excitation Laser
Red Laser (633 nm)
Application Notes

Additional reported application (for relevant formats) include: spatial biology (IBEX)1,2.

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)
  1. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  2. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Santini-González J, et al. 2022. Front Endocrinol (Lausanne). 13:989815. PubMed
  2. Sun L, et al. 2021. Cancer Cell. 39:1361. PubMed
  3. Kang YA, et al. 2023. J Exp Med. 220:. PubMed
  4. Koop K, et al. 2023. Front Immunol. 14:1163198. PubMed
  5. Lin YN, et al. 2022. Oncoimmunology. 11:2027136. PubMed
  6. Otano I, et al. 2021. Nat Commun. 12:7296. PubMed
  7. Hiraoka N, et al. 2011. Gastroenterology. 140:310. PubMed
  8. Baptista AP et al. 2019. Immunity. 50(5):1188-1201 . PubMed
  9. Roufaiel M, et al. 2016. Nat Immunol. 10.1038/ni.3564. PubMed
  10. Webster HC, et al. 2020. J Immunol Methods. 112702:477. PubMed
  11. Rodriguez AB, et al. 2021. Cell Reports. 36(3):109422. PubMed
  12. Etxeberria I, et al. 2020. Cancer Cell. 36(6):613-629. PubMed
  13. Zeng Z, et al. 2022. Oncogene. :. PubMed
  14. Guldner IH, et al. 2020. Cell. 183(5):1234-1248.e25. PubMed
  15. Rassy D, et al. 2020. J Neuropathol Exp Neurol. 226:79. PubMed
  16. Xiong S 2013. J Immunol. 190:3267. PubMed
  17. Robertson TF, et al. 2021. J Cell Biol. 220:. PubMed
  18. Sluis T, et al. 2015. Clin Cancer Res. 21:781. PubMed
  19. Luck H, et al. 2019. Nat Commun. 10:3650. PubMed
  20. Cecchinato V, et al. 2017. J Immunol. 198(1):184-195. PubMed
  21. Chakraborty M, et al. 2021. Cell Reports. 34(2):108609. PubMed
  22. Zhang H, et al. 2021. Cell Reports. 35(6):109096. PubMed
  23. Ovadya Y, et al. 2018. Nat Commun. 9:5435. PubMed
  24. Ardehali R, et al. 2013. Proc Natl Acad Sci U S A. 110:3405. PubMed
  25. Agudo J et al. 2018. Immunity. 48(2):271-285 . PubMed
  26. Smith KJ, et al. 2022. PLoS Biol. 20:e3001554. PubMed
  27. Chen S, et al. 2015. Blood. 126: 103 - 112. PubMed
  28. Crinier A, et al. 2018. Immunity. 49:971. PubMed
  29. Bárcena C et al. 2019. EBioMedicine. 43:513-524 . PubMed
  30. Shi H et al. 2018. Immunity. 49(5):899-914 . PubMed
  31. Chang H, et al. 2016. J Immunol. 197: 2473 - 2484. PubMed
  32. Lerbs T, et al. 2020. JCI Insight. 5:00. PubMed
  33. O'Reilly LA, et al. 2018. Immunity. 48:570. PubMed
RRID
AB_389323 (BioLegend Cat. No. 100209)

Antigen Details

Structure
Ig superfamily, CD3/TCR, 20 kD
Distribution

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

Function
Antigen recognition, TCR signal transduction, T cell activation
Ligand/Receptor
Peptide antigen/MHC-complex
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
12502 View all products for this Gene ID
UniProt
View information about CD3 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All CD3 Reagents Request Custom Conjugation
Description Clone Applications
FITC anti-mouse CD3 17A2 FC
PE anti-mouse CD3 17A2 FC
Purified anti-mouse CD3 17A2 FC,IHC-F,IP,ICC
Alexa Fluor® 647 anti-mouse CD3 17A2 FC,IHC-F,3D IHC,SB
Alexa Fluor® 488 anti-mouse CD3 17A2 FC,IHC-F,3D IHC
Pacific Blue™ anti-mouse CD3 17A2 FC
Alexa Fluor® 700 anti-mouse CD3 17A2 FC
PerCP/Cyanine5.5 anti-mouse CD3 17A2 FC
PE/Cyanine7 anti-mouse CD3 17A2 FC
APC/Cyanine7 anti-mouse CD3 17A2 FC
Brilliant Violet 421™ anti-mouse CD3 17A2 FC,ICC
Brilliant Violet 570™ anti-mouse CD3 17A2 FC
Brilliant Violet 650™ anti-mouse CD3 17A2 FC
Brilliant Violet 785™ anti-mouse CD3 17A2 FC
Brilliant Violet 510™ anti-mouse CD3 17A2 FC
APC anti-mouse CD3 17A2 FC
Ultra-LEAF™ Purified anti-mouse CD3 17A2 FC,IHC-F,IP,ICC
Brilliant Violet 605™ anti-mouse CD3 17A2 FC
Alexa Fluor® 594 anti-mouse CD3 17A2 IHC-F,FC,3D IHC,SB
Brilliant Violet 711™ anti-mouse CD3 17A2 FC
Biotin anti-mouse CD3 17A2 FC,IHC-F
PE/Dazzle™ 594 anti-mouse CD3 17A2 FC
APC/Fire™ 750 anti-mouse CD3 17A2 FC
Brilliant Violet 750™ anti-mouse CD3 17A2 FC
TotalSeq™-A0182 anti-mouse CD3 17A2 PG
TotalSeq™-B0182 anti-mouse CD3 17A2 PG
Spark Blue™ 550 anti-mouse CD3 17A2 FC
Spark NIR™ 685 anti-mouse CD3 17A2 FC
TotalSeq™-C0182 anti-mouse CD3 17A2 PG
APC/Fire™ 810 anti-mouse CD3 17A2 FC
PE/Fire™ 640 anti-mouse CD3 17A2 FC
Spark YG™ 570 anti-mouse CD3 17A2 IHC-F
PE/Fire™ 700 anti-mouse CD3 17A2 FC
PE/Cyanine5 anti-mouse CD3 17A2 FC
Spark Blue™ 574 anti-mouse CD3 Antibody 17A2 FC
Spark Violet™ 423 anti-mouse CD3 17A2 FC
PE/Fire™ 810 anti-mouse CD3 17A2 FC
Spark Red™ 718 anti-mouse CD3 17A2 FC
Spark UV™ 387 anti-mouse CD3 17A2 FC
Go To Top Version: 8    Revision Date: 01.24.2024

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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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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