Left: Staining of mouse splenocytes with PE-Cy7 anti-mouse CD25 (PC61.5) (cat. 25-0251) and 0.25 ĩg of AlexaFluor647 anti-mouse FR4. Right: analysis of gated CD4+CD25+ cells stained with AlexaFluor 647 isotype control (cat. 51-4031)(open) or AlexaFluor647 anti-mouse FR4 (colored).
Contents: Alexa FluorŽ 647 anti-mouse FR4 (Folate receptor 4, FBP, FRδ) Catalog Number: 51-5445 Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN3, BSA and/or gelatin for protein stabilization Clone: eBio12A5 Host/Isotype: Rat IgG2b, κ Storage Conditions: Store at 4°C. DO NOT FREEZE. LIGHT-SENSITIVE MATERIAL.
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51-5445-80
25 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.
Description
The monoclonal antibody eBio12A5 recognizes FR4, also known as Folate receptor 4, FR δ and FBP3 (folate binding protein3). FR4 is a heavily glycosylated 35 kD receptor for folic acid and the physiologic circulating form of the vitamin, N5-methyltetrahydrofolate. Natural T regs have high levels of FR4 and together with CD25 can be used to distinguish natural Tregs, effector T cells, memory-like T cells and naīve T cells. FR4hi CD25hi cells are natural Tregs with high levels of Foxp3. FR4hi CD25lo cells are identified as central memory T cells which upon stimulation can proliferate and produce large amounts of IL-2. In contrast FR4lo CD25hi secrete proinflammatory cytokines such as IFNγ and IL-17 and have been characterized as effector memory T cells.
Based on co-staining studies, the epitopes recognized by eBioTH6 (cat 51-5446) and eBio12A5 are different thereby allowing functional studies with eBioTH6 to be evaluated with eBio12A5.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This eBio12A5 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This eBio12A5 antibody has been tested by flow cytometric analysis of mouse splenocytes. 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
Elnakat H, Ratnam M. Distribution, functionality and gene regulation of folate receptor isoforms: implications in targeted therapy.
Adv Drug Deliv Rev. 2004 Apr 29;56(8):1067-84.
Spiegelstein O, Eudy JD, Finnell RH. Identification of two putative novel folate receptor genes in humans and mouse.
Gene. 2000 Nov 27;258(1-2):117-25.
Sugimoto N, Oida T, Hirota K, Nakamura K, Nomura T, Uchiyama T, Sakaguchi S. Foxp3-dependent and -independent molecules specific for CD25+CD4+ natural regulatory T cells revealed by DNA microarray analysis.
Int Immunol. 2006 Aug;18(8):1197-209.
Yamaguchi T, Hirota K, Nagahama K, Ohkawa K, Takahashi T, Nomura T, Sakaguchi S. Control of immune response by antigen-specific regulatory T cells expressing the folate receptor. Immunity July 2007 ePub (eBio12A5 FC, IP)
Legal
Alexa FluorŽ and Pacific BlueŽ 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.
