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Alexa Fluor® 647 anti-human CD25 Antibody

Pricing & Availability
Clone
M-A251 (See other available formats)
Regulatory Status
RUO
Workshop
IV A053
Other Names
IL-2 receptor α chain, Low affinity IL-2R, IL-2Rα chain
Isotype
Mouse IgG1, κ
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Product Citations
publications
1_M-A251_AF647_CD25_Antibody_FC_051414
PHA-stimulated (3 days) human peripheral blood lymphocytes were stained with CD25 (clone M-A251) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 1_M-A251_AF647_CD25_Antibody_FC_051414
    PHA-stimulated (3 days) human peripheral blood lymphocytes were stained with CD25 (clone M-A251) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 2_8_Human_LN_CD1c_CD8_CD25
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD1c (red) in Cycle 3, CD8 (blue) in Cycle 4, and CD25 (yellow) 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).
Compare all formats See Alexa Fluor® 647 spectral data
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356127 25 tests $124
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356128 100 tests $265
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Description

CD25 is a 55 kD type I transmembrane glycoprotein also known as low affinity IL-2 receptor α chain or Tac. It is expressed on progenitor lymphocytes, activated T and B cells, and activated monocytes/macrophages. CD25 is also expressed on a subset of non-stimulated CD4+ T cells termed T regulatory cells. Soluble CD25/IL-2Rα is produced as a consequence of lymphocyte stimulation and is found in biological fluids following inflammatory responses. CD25 associates with IL-2 receptor β (CD122) and common γ (CD132) chains to form a high affinity IL-2R complex.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Reported Reactivity
Baboon, Cynomolgus, Rhesus
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Human PHA-induced lymphocyte cells
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

Additional reported applications (for the relevant formats) include: immunohistochemical staining of paraformaldehyde fixed frozen sections1 and spatial biology (IBEX)2,3.

The CD25 molecule reveals three epitope regions: A, B, and C. M-A251 antibody recognizes epitope region B. Unlike other CD25 antibody clones, M-A251 can detect CD25 after fixation with paraformaldehyde.

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

(PubMed link indicates BioLegend citation)
  1. Li H and Pauza CD. 2015. Eur. J. Immunol. 45:298. (IHC)
  2. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  3. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Thaker YR, et al. 2022. Front Oncol. 12:884196. PubMed
  2. Wang L, et al. 2021. Vaccines (Basel). :9. PubMed
RRID
AB_2563587 (BioLegend Cat. No. 356127)
AB_2563588 (BioLegend Cat. No. 356128)

Antigen Details

Structure
Type I transmembrane glycoprotein, 55 kD; low-affinity IL-2 receptor α chain
Distribution

Activated T and B cells, monocytes/macrophages, Tregs

Interaction
Associates with IL-2Rβ/CD122 and IL-2Rγ/CD132 receptor chains to form a high-affinity IL-2R complex
Ligand/Receptor
IL-2
Cell Type
B cells, Macrophages, Monocytes, T cells, Tregs
Biology Area
Cell Biology, Immunology, Neuroscience, Neuroscience Cell Markers
Molecular Family
CD Molecules, Cytokine/Chemokine Receptors
Antigen References

1. Knapp W, et al. 1989. Leucocyte Typing IV: White Cell Differentiation Antigens. Oxford University Press.
2. Schlossman S, et al. 1995. Leucocyte Typing V: White Cell Differentiation Antigens. Oxford University Press.
3. Barclay N, et al. 1997. The Leukocyte Antigen FactsBook. Academic Press Inc.
4. Taniguchi T and Minami Y. et al. 1993. Cell 73:5.
5. Waldmann T. 1991. J. Biol. Chem. 266:2681.

Gene ID
3559 View all products for this Gene ID
UniProt
View information about CD25 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 CD25 Reagents Request Custom Conjugation
Description Clone Applications
APC/Cyanine7 anti-human CD25 M-A251 FC
Purified anti-human CD25 M-A251 FC,IHC-F
PE anti-human CD25 M-A251 FC
FITC anti-human CD25 M-A251 FC
PE/Cyanine7 anti-human CD25 M-A251 FC
APC anti-human CD25 M-A251 FC
PerCP/Cyanine5.5 anti-human CD25 M-A251 FC
Brilliant Violet 421™ anti-human CD25 M-A251 FC
Alexa Fluor® 488 anti-human CD25 M-A251 FC
Alexa Fluor® 700 anti-human CD25 M-A251 FC
Brilliant Violet 510™ anti-human CD25 M-A251 FC
PE/Dazzle™ 594 anti-human CD25 M-A251 FC
Biotin anti-human CD25 M-A251 FC
Alexa Fluor® 647 anti-human CD25 M-A251 FC,SB
Pacific Blue™ anti-human CD25 M-A251 FC
PerCP anti-human CD25 M-A251 FC
APC/Fire™ 750 anti-human CD25 M-A251 FC
Brilliant Violet 711™ anti-human CD25 M-A251 FC
Brilliant Violet 785™ anti-human CD25 M-A251 FC
Brilliant Violet 605™ anti-human CD25 M-A251 FC
KIRAVIA Blue 520™ anti-human CD25 M-A251 FC
PE/Fire™ 700 anti-human CD25 M-A251 FC
APC/Fire™ 810 anti-human CD25 M-A251 FC
Spark NIR™ 685 anti-human CD25 Antibody M-A251 FC
Spark YG™ 581 anti-human CD25 M-A251 FC
PE/Fire™ 640 anti-human CD25 Antibody M-A251 FC
PE anti-human CD25 M-A251 FC
PerCP/Cyanine5.5 anti-human CD25 M-A251 FC
APC/Fire™ 750 anti-human CD25 M-A251 FC
PE/Cyanine7 anti-human CD25 M-A251 FC
APC anti-human CD25 M-A251 FC
PE/Cyanine5 anti-human CD25 M-A251 FC
FITC anti-human CD25 M-A251 FC
Spark Red™ 718 anti-human CD25 M-A251 FC
GMP PE anti-human CD25 M-A251 FC
GMP PE/Cyanine7 anti-human CD25 M-A251 FC
PerCP/Fire™ 780 anti-human CD25 M-A251 FC
PE/Fire™ 744 anti-human CD25 M-A251 FC
PerCP/Fire™ 806 anti-human CD25 M-A251 FC
Brilliant Violet 650™ anti-human CD25 M-A251 FC
Go To Top Version: 5    Revision Date: 04/22/2022

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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