Spark Blue™ 574 anti-mouse CD25 (Flexi-Fluor™) Fluor Antibody, CD25, PC61

Clone: PC61 (See other available formats)
Regulatory Status: RUO
Other Names: IL-2Rα, Ly-43, p55, Tac
Isotype: Rat IgG1, λ
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Product Citations: publications

Compare all formats See Spark Blue™ 574 spectral data

Cat #	Size	Price	Save
285179	50 µg	¥73,920

Description

CD25 is a 55 kD glycoprotein also known as the low affinity IL-2Rα, Ly-43, p55, or Tac. It is expressed on activated T and B cells, thymocyte subsets, pre-B cells, and T regulatory cells. In association with CD122 (IL-2Rβ) and CD132 (common γ chain), CD25 forms the high affinity signaling IL-2 receptor.

Product Details

Technical data sheet

Product Details

Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: IL-2-dependent cytolytic mouse T-cell clone B6.1
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 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.
Excitation Laser: Blue Laser (488 nm)
Application Notes: Additional reported applications (for the relevant formats) include: immunoprecipitation^1,2, in vitro blocking of IL-2 binding to low- and high-affinity receptors^1-4, growth inhibition of IL-2-dependent T-cell lines^1-4, in vivo depletion of CD25⁺CD4⁺ Treg cells^5-8,10, and immunohistochemical staining of acetone-fixed frozen sections². PC61 antibody recognizes a different epitope than 3C7 antibody (Cat. No. 101902). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 102040) with endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered.
Additional Product Notes: For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.
RRID: AB_3662519 (BioLegend Cat. No. 285179)

Antigen Details

Structure: Forms high affinity IL-2R with IL-2Rβ (CD122) and IL-2Rγ (CD132), 55 kD
Distribution: Activated T cells and B cells, thymocyte subset, pre-B cells, T regulatory cells
Function: IL-2 receptor
Ligand/Receptor: IL-2
Cell Type: B cells, T cells, Thymocytes, Tregs
Biology Area: Immunology
Molecular Family: CD Molecules, Cytokine/Chemokine Receptors
Antigen References: Taniguchi T, et al. 1993. Cell 73:5-8.

Waldmann TA. 1991. J Biol Chem. 266:2681-4.

Read S, et al. 2000. J Exp Med. 192:295-302.

Lowenthal JW, et al. 1985. J Immunol. 135:3988-94.
Gene ID: 16184 View all products for this Gene ID
UniProt: View information about CD25 on UniProt.org

Documentation

Reviews

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Related Protocols

Cell Surface Flow Cytometry Staining Protocol

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Related Pages & Pathways

Pathways

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.

Other Formats

View All CD25 Reagents Request Custom Conjugation

Description	Clone	Applications
APC anti-mouse CD25	PC61	FC
Biotin anti-mouse CD25	PC61	FC
FITC anti-mouse CD25	PC61	FC
PE anti-mouse CD25	PC61	FC
PE/Cyanine5 anti-mouse CD25	PC61	FC
Purified anti-mouse CD25	PC61	FC,IHC-F,IP,Block,Depletion
PE/Cyanine7 anti-mouse CD25	PC61	FC
Alexa Fluor® 488 anti-mouse CD25	PC61	FC
Alexa Fluor® 647 anti-mouse CD25	PC61	FC
Pacific Blue™ anti-mouse CD25	PC61	FC
Alexa Fluor® 700 anti-mouse CD25	PC61	FC
APC/Cyanine7 anti-mouse CD25	PC61	FC
PerCP/Cyanine5.5 anti-mouse CD25	PC61	FC
PerCP anti-mouse CD25	PC61	FC
Brilliant Violet 421™ anti-mouse CD25	PC61	FC
Brilliant Violet 605™ anti-mouse CD25	PC61	FC
Brilliant Violet 650™ anti-mouse CD25	PC61	FC
Ultra-LEAF™ Purified anti-mouse CD25	PC61	FC,IHC-F,IP,Block,Depletion
Brilliant Violet 510™ anti-mouse CD25	PC61	FC
PE/Dazzle™ 594 anti-mouse CD25	PC61	FC
Brilliant Violet 711™ anti-mouse CD25	PC61	FC
Brilliant Violet 785™ anti-mouse CD25	PC61	FC
Alexa Fluor® 594 anti-mouse CD25	PC61	IHC-F
APC/Fire™ 750 anti-mouse CD25	PC61	FC
TotalSeq™-A0097 anti-mouse CD25	PC61	PG
KIRAVIA Blue 520™ anti-mouse CD25	PC61	FC
TotalSeq™-B0097 anti-mouse CD25	PC61	PG
TotalSeq™-C0097 anti-mouse CD25	PC61	PG
Spark NIR™ 685 anti-mouse CD25 Antibody	PC61	FC
PE/Fire™ 640 anti-mouse CD25	PC61	FC
Spark YG™ 581 anti-mouse CD25	PC61	FC
APC/Fire™ 810 anti-mouse CD25	PC61	FC
Brilliant Violet 750™ anti-mouse CD25	PC61	FC
PerCP/Fire™ 780 anti-mouse CD25	PC61	FC
PE/Fire™ 700 anti-mouse CD25	PC61	FC
Spark PLUS UV395™ anti-mouse CD25	PC61	FC
Spark Blue™ 574 anti-mouse CD25 (Flexi-Fluor™)	PC61	FC

Certificate of Analysis

Go To Top Version: 1 Revision Date: 09/06/2024

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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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.