Spark YG™ 593 anti-human CD25 (Flexi-Fluor™) 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
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285447 50 µg 316€
 
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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 (pdf)

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
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark YG™ 593 under optimal conditions.
Concentration
0.2 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

Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

* Spark YG™ 593 has a maximum excitation of 573 nm and a maximum emission of 593 nm.

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

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References
  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

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

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool

What if my requested antibody-dye combination doesn't perform as expected?

Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.

Other Formats

View All CD25 Reagents Request Custom Conjugation
Description Clone Applications
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
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
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
PerCP/Fire™ 806 anti-human CD25 M-A251 FC
PE/Fire™ 744 anti-human CD25 M-A251 FC
Brilliant Violet 650™ anti-human CD25 M-A251 FC
Brilliant Violet 750™ anti-human CD25 M-A251 FC
StarBright UltraViolet 740 anti-human CD25 M-A251 FC
Spark YG™ 593 anti-human CD25 (Flexi-Fluor™) Antibody M-A251 FC
Spark PLUS B574™ anti-human CD25 Antibody M-A251 FC
Go To Top Version: 1    Revision Date: 06/02/2025

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.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
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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