Alexa Fluor® 647 anti-human TCR Vα7.2 Antibody

Pricing & Availability
Clone
3C10 (See other available formats)
Regulatory Status
RUO
Other Names
T cell antigen receptor Vα7.2
Isotype
Mouse IgG1, κ
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Product Citations
publications
3C10_AF647_TCRVa7.2_Antibody_FC_1_092315
Human peripheral blood lymphocytes were stained with CD3 PE and TCR Vα7.2 (clone 3C10) Alexa Fluor® 647 (top), or mouse IgG1, κ Alexa Fluor® 647 isotype control (bottom).
  • 3C10_AF647_TCRVa7.2_Antibody_FC_1_092315
    Human peripheral blood lymphocytes were stained with CD3 PE and TCR Vα7.2 (clone 3C10) Alexa Fluor® 647 (top), or mouse IgG1, κ Alexa Fluor® 647 isotype control (bottom).
  • 3C10_AF647_TCRVa7.2_Antibody_FC_2_102315
  • 9_Human_LN_HLA-DR_Va72
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: HLA-DR (purple) in Cycle 3 and Vα7.2 (yellow) in Cycle 7. 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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351725 25 tests 128€
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351726 100 tests 296€
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Description

The 3C10 antibody recognizes the Vα7.2 T cell antigen receptor (TCR) α-chain segment which, joined with the Jα33 segment, constitutes an invariant TCR that is a characteristic of the mucosal-associated invariant T cells (MAIT cells). MAIT cells are restricted by a nonpolymorphic class Ib major histocompatibility complex (MHC) molecule, MHC-related molecule 1 (MR1). MAIT cells are present in human blood (1-8% of T cells), mesenteric lymph nodes, liver, and intestinal mucosa.  MAIT cells play a role in detecting and fighting off microbial infections.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant TCR
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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
SB - Reported in the literature, not verified in house

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 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

* 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

Associated with an anti-CD161 or -IL18Ra staining, the 3C10 antibody allows unequivocal identification of MAIT cells. Importantly, the Va7.2  segment can also be used by conventional T cells. Therefore, the 3C10 also stains a subset of conventional CD4 and CD8 T cells. Additional reported applications (for the relevant formats) include: spatial biology (IBEX)3,4.

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).

Application References
  1. Martin E, et al. 2009. PLoS Biol. 7:525.
  2. Wakao H, et al. 2013. Cell Stem Cell 12:1. PubMed
  3. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  4. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID
AB_2566334 (BioLegend Cat. No. 351725)
AB_2566334 (BioLegend Cat. No. 351726)

Antigen Details

Structure
Invariant T cell antigen receptor (TCR) α-chain (iTCRa), composed of the invariant α-chain variable region 7.2 (iVα7.2) segment in humans
Distribution

Mucosal-associated invariant T (MAIT) cells and conventional T cells

Cell Type
T cells
Biology Area
Adaptive Immunity, Immunology
Molecular Family
TCRs
Antigen References

1. Le Bourhis L, et al. 2010. Nat. Immunol. 11:701.

Gene ID
6955 View all products for this Gene ID
UniProt
View information about TCR Valpha7.2 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.

Go To Top Version: 2    Revision Date: 04/18/2022

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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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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