Product Information

Contents: Allophycocyanin-Alexa Fluor® 750 (APC-Alexa Fluor® 750) anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55)
Catalog Number: 27-0251
Sizes: 50 ug, 100 ug
Formulation: Phosphate buffer pH 7.2,
500 mM NaCl, 0.09% NaN₃, 0.1% gelatin
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
This tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling & experimental procedures. This product is guaranteed for 6 months upon receipt when stored properly.
Clone: PC61.5
Isotype: Rat IgG1, λ

Staining of 3-day ConA activated C57Bl/6 splenocytes with staining buffer (autofluorescence) (left) or 0.25 μg of APC-Alexa Fluor® 750 anti-mouse CD25 (PC61.5) (right). Cells in the lymphocyte gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
11-0251	May be discontinued; see superior product Alexa Fluor® 488 FITC anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	488	518	FC
12-0251	PE anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	488	575	FC
13-0251	Biotin anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	N/A	N/A	FC
14-0251	Affinity Purified anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	N/A	N/A	FC IHC IP
15-0251	PE-Cy5 anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	488	670	FC
16-0251	Functional Grade* Purified anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	N/A	N/A	FA FC
17-0251	APC anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	633	660	FC
25-0251	Phycoerythrin-Cy7 (PE-Cy7) anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	488	760	FC
27-0251	Allophycocyanin-Alexa Fluor® 750 (APC-Alexa Fluor® 750) anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55) (APC-Cy7 Equivalent)	633	779	FC
36-0251	Functional Grade* Biotin anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	N/A	N/A	FC
45-0251	PerCP-Cy5.5 anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55)	488	690	FC
53-0251	Alexa Fluor® 488 anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra)	488	519	FC
56-0251	Alexa Fluor® 700 anti-mouse CD25 (Interleukin-2 Receptor alpha, IL-2 Receptor alpha, IL-2Ra, p55, IL2Ra) (APC-Cy5.5 Equivalent)	633	723	FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The PC61.5 antibody reacts with mouse CD25, the 55 kDa interleukin-2 receptor α chain (IL-2Rα). CD25 is expressed by early progenitors of the T and B lineage as well as by activated mature T and B lymphocytes. By itself, CD25 binds IL-2 only with low affinity. However, CD25 associates with CD122 (IL-2 receptor beta chain) and CD132 (common γ chain) to form the high affinity IL-2 receptor. Binding of IL-2 to both the high and low affinity classes of IL-2 receptor is inhibited by the PC61.5 antibody. CD25 plays a role in lymphocyte differentiation and activation/proliferation.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This PC61.5 antibody has been reported for use in flow cytometric analysis.

The photo-emission properties and compensation required for Alexa Fluor® 750 are similar to APC-Cy7; however Alexa Fluor® 750 has several advantages when compared to APC-Cy7. The Alexa Fluor® 750 has peak emission of 779 nm, and usually has a slightly higher mean fluorescent intensity when compared to the same clone conjugated with APC-Cy7, which has peak emission of 767 nm. Furthermore, Alexa Fluor® 750 is considered to be a more photostable fluorochrome as compared with APC-Cy7. It is advised not to use fixatives with Alexa Fluor® 750 as this may lead to degradation of the fluorochrome and loss of staining.

Applications Tested

This PC61.5 antibody has been tested by flow cytometric analysis of mouse splenocyte cell suspensions and ConA-activated splenocyte culture. This can be used at less than or equal to 0.5 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Lowenthal, J. W., P. Corthesy, et al. (1985)."High and low affinity IL 2 receptors: analysis by IL 2 dissociation rate and reactivity with monoclonal anti-receptor antibody PC61." J Immunol 135(6): 3988-94.

Lowenthal, J. W., C. Tougne, et al. (1985). "Antigenic stimulation regulates the expression of IL 2 receptors in a cytolytic T lymphocyte clone." J Immunol 134(2): 931-9.

Lowenthal, J. W., R. H. Zubler, et al. (1985)."Similarities between interleukin-2 receptor number and affinity on activated B and T lymphocytes." Nature 315(6021): 669-72.

Hayashi, T, Hasegawa, K, Adachi, C. (2005) Elimination of CD4+CD25+ T cell accelerates the development of glomerulonephritis during the preactive phase in autoimmune-prone female NZB × NZW F1 mice. Int J Exp Path 86: 289-296 (PC61.5, in vivo depletion, PubMed)

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Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.