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Alexa Fluor® 594 anti-human CD3 Antibody

Pricing & Availability
Clone
UCHT1 (See other available formats)
Regulatory Status
RUO
Workshop
III 471
Other Names
T3, CD3ε
Isotype
Mouse IgG1, κ
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Product Citations
publications
1_UCHT1_A594_CD3_Antibody_1_102820
Human peripheral blood mononuclear cells were fixed with 2% paraformaldehyde (PFA), and then stained with 10 µg/ml CD14 (clone HCD14) Alexa Fluor® 647 (yellow), 10 µg/ml CD19 (clone HIB19) Alexa Fluor® 488 (green), and 10 µg/ml CD3 (clone UCHT1) Alexa Fluor® 594 (red) for 30 minutes at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 40X objective.
  • 1_UCHT1_A594_CD3_Antibody_1_102820
    Human peripheral blood mononuclear cells were fixed with 2% paraformaldehyde (PFA), and then stained with 10 µg/ml CD14 (clone HCD14) Alexa Fluor® 647 (yellow), 10 µg/ml CD19 (clone HIB19) Alexa Fluor® 488 (green), and 10 µg/ml CD3 (clone UCHT1) Alexa Fluor® 594 (red) for 30 minutes at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 40X objective.
  • 2_UCHT1_AF594_FC_111214
    Human peripheral blood lymphocytes were stained with CD3 (clone UCHT1) Alexa Flour® 594 (filled histogram) or mouse IgG1, κ Alexa Flour® 594 isotype control (open histogram). The data was acquired by BD LSRFortessa™ cell analyzer equipped with the Yellow-Green Laser (561 nm).
  • 3_1_Human_LN_BCL2_CD3
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: BCL2 (cyan) in Cycle 1, CD3 (purple) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 4_12_Human_LN_CD3_DC-SIGN
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD3 (purple) in Cycle 1 and DC-SIGN (cyan) in Cycle 9. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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300446 100 µg £182
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Description

CD3ε is a 20 kD chain of the CD3/T-cell receptor (TCR) complex which is composed of two CD3ε, one CD3γ, one CD3δ, one CD3ζ (CD247), and a T-cell receptor (α/β or γ/δ) heterodimer. It is found on all mature T cells, NKT cells, and some thymocytes. CD3, also known as T3, is a member of the immunoglobulin superfamily that plays a role in antigen recognition, signal transduction, and T cell activation.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Reported Reactivity
Chimpanzee
Antibody Type
Monoclonal
Host Species
Mouse
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® 594 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

ICC - Quality tested
FC - Verified
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 2.5 - 10 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


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

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Excitation Laser
Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections4,6,7 and formalin-fixed paraffin-embedded sections11, immunoprecipitation1, activation of T cells2,3,5, Western blotting9, and spatial biology (IBEX)16,17. The LEAF™ purified antibody (Endotoxin < 0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 300413, 300414, and 300432). For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 300437, 300438, 300465, 300466, 300473, 300474) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin < 0.01 EU/µg).

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

View more applications data for this product in our Scientific Poster Library.

Application References

(PubMed link indicates BioLegend citation)
  1. Salmeron A, et al. 1991. J. Immunol. 147:3047. (IP)
  2. Graves J, et al. 1991. J. Immunol. 146:2102. (Activ)
  3. Lafont V, et al. 2000. J. Biol. Chem. 275:19282. (Activ)
  4. Ryschich E, et al. 2003. Tissue Antigens 62:48. (IHC)
  5. Thompson AG, et al. 2004. J. Immunol. 173:1671. (Activ)
  6. Sakkas LI, et al. 1998. Clin. Diagn. Lab. Immun. 5:430. (IHC)
  7. Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC)
  8. Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed
  9. Van Dongen JJM, et al. 1988. Blood 71:603. (WB)
  10. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  11. Pollard, K. et al. 1987. J. Histochem. Cytochem. 35:1329. (IHC)
  12. Luckashenak N, et al. 2013. J. Immunol. 190:27. PubMed
  13. Laurent AJ, et al. 2014. PLoS One. 9:103683. PubMed
  14. Li J, et al. 2015. Cancer Res. 75:508. PubMed
  15. Stoeckius M, et al. 2017. Nat. Methods. 14:865-868. (PG)
  16. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  17. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Emmons TR, et al. 2021. Cancer Immunol Res. 9:790. PubMed
  2. Mehta AK, et al. 2021. Nat Cancer. 2:66. PubMed
  3. Su Y, et al. 2021. Cancers (Basel). 13:. PubMed
RRID
AB_2563236 (BioLegend Cat. No. 300446)

Antigen Details

Structure
Ig superfamily, with the subunits of CD3γ, CD3δ, CD3ζ (CD247) and TCR (α/β or γ/δ) forms CD3/TCR complex, 20 kD
Distribution

Mature T and NK T cells, thymocyte differentiation

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

1. Barclay N, et al. 1993. The Leucocyte FactsBook. Academic Press. San Diego.
2. Beverly P, et al. 1981. Eur. J. Immunol. 11:329.
3. Lanier L, et al. 1986. J. Immunol. 137:2501-2507.

Gene ID
916 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
APC anti-human CD3 UCHT1 FC
Biotin anti-human CD3 UCHT1 FC
FITC anti-human CD3 UCHT1 FC
PE anti-human CD3 UCHT1 FC
PE/Cyanine5 anti-human CD3 UCHT1 FC
Purified anti-human CD3 UCHT1 FC,CyTOF®,IHC-F,IP,Activ,WB
Alexa Fluor® 647 anti-human CD3 UCHT1 FC,ICC,IHC-F
Alexa Fluor® 488 anti-human CD3 UCHT1 FC,ICC,IHC-F
Pacific Blue™ anti-human CD3 UCHT1 FC
PE/Cyanine7 anti-human CD3 UCHT1 FC
Alexa Fluor® 700 anti-human CD3 UCHT1 FC
APC/Cyanine7 anti-human CD3 UCHT1 FC
PerCP anti-human CD3 UCHT1 FC
PerCP/Cyanine5.5 anti-human CD3 UCHT1 FC
Brilliant Violet 421™ anti-human CD3 UCHT1 FC,ICC,IHC-F
Brilliant Violet 570™ anti-human CD3 UCHT1 FC
Ultra-LEAF™ Purified anti-human CD3 UCHT1 FC,CyTOF®,IHC-F,IP,Activ,WB
Purified anti-human CD3 (Maxpar® Ready) UCHT1 FC,CyTOF®
Alexa Fluor® 594 anti-human CD3 UCHT1 ICC,FC,SB
PE/Dazzle™ 594 anti-human CD3 UCHT1 FC
Brilliant Violet 510™ anti-human CD3 UCHT1 FC
Brilliant Violet 605™ anti-human CD3 UCHT1 FC
Brilliant Violet 711™ anti-human CD3 UCHT1 FC
Brilliant Violet 650™ anti-human CD3 UCHT1 FC
APC/Fire™ 750 anti-human CD3 UCHT1 FC
Pacific Blue™ anti-human CD3 UCHT1 FC
Brilliant Violet 785™ anti-human CD3 UCHT1 FC
PE/Dazzle™ 594 anti-human CD3 UCHT1 FC
TotalSeq™-A0034 anti-human CD3 UCHT1 PG
TotalSeq™-B0034 anti-human CD3 UCHT1 PG
TotalSeq™-C0034 anti-human CD3 UCHT1 PG
PE anti-human CD3 UCHT1 FC
PE/Cyanine7 anti-human CD3 UCHT1 FC
KIRAVIA Blue 520™ anti-human CD3 UCHT1 FC
Spark Violet™ 538 anti-human CD3 Antibody UCHT1 FC
TotalSeq™-D0034 anti-human CD3 UCHT1 PG
Spark Blue™ 574 anti-human CD3 Antibody UCHT1 FC
GMP Pacific Blue™ anti-human CD3 UCHT1 FC
GMP PE anti-human CD3 UCHT1 FC
GMP PE/Dazzle™ 594 anti-human CD3 UCHT1 FC
Spark Violet™ 423 anti-human CD3 UCHT1 FC
GMP PE/Cyanine7 anti-human CD3 UCHT1 FC
Spark Blue™ 515 anti-human CD3 UCHT1 FC
Go To Top Version: 3    Revision Date: 04/26/2022

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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