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Alexa Fluor® 647 anti-human FOXP3 Flow Kit；Clone#：206D；Host/Isotype：Mouse IgG1, κ   

详细描述

Clone: 206D
Isotype: Mouse IgG1, κ
Reactivity: Human, Cross-Reactivity: Baboon, Cynomolgus, Rhesus, Pigtailed Macaque
Storage & Handling: This kit is guaranteed for three months. Upon receipt, store between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

ICFC - Quality tested
Application Notes:

Materials Provided:
1. Alexa Fluor® 647 anti-human FOXP3
25 tests
2. Alexa Fluor® 647 Mouse IgG1, k isotype control
25 tests
3. FOXP3 Fix/Perm buffer set, 100 tests

Materials not included:
Cell Staining Buffer
anti-human CD4 PE-Cy5/CD25 PE cocktail

Surface Staining & FOXP3 Buffer Preparation:

Centrifugation steps: perform at 250Xg for 5min
Incubation steps: perform at room temperature

1. Perform cell surface staining if necessary.
2. Prepare 1X buffer solutions of FOXP3 Fix/Perm buffer and FOXP3 Perm buffer in PBS.

NOTE: The FOXP3 Perm buffer (10X) may have crystalization or precipitation observed when it is stored at 2-8°C, however, it is normal and does not affect the buffer performance. If there is a heavy precipitation observed after diluting to 1X working solution, it may be clarified by filtering.
Caution: The FOXP3 Fix/Perm buffer contains paraformaldehyde, which is toxigenic and mutagenic. Please handle with caution and wear gloves, lab coat and necessary protection to avoid direct body contact.

FOXP3 Intracellular Staining Procedures:

3. Add 1 ml of 1X FOXP3 Fix/Perm solution to each tube, resuspend the cells (gentle vortex) and incubate at room temperature in the dark for 20 minutes, then centrifuge and remove the supernatant. The cell pellet will now be translucent and difficult to see; take care not to dislodge and accidentally aspirate cells at all later stages of staining protocol.
4. Wash: resuspend cells in cell staining buffer; centrifuge, then discard the supernatant.
5. Wash: resuspend in 1ml 1X FOXP3 Perm buffer; centrifuge, then discard the supernatant.
6. Resuspend cells in 1ml 1X FOXP3 Perm buffer, incubate in the dark for 15 minutes; centrifuge, then discard the supernatant. Resuspend the pellet in 100 µl of 1X FOXP3 Perm buffer.
7. Add appropriate amount of flurochrome conjugated anti-FOXP3 antibody and incubate at room temperature in the dark for 30 minutes.
8. Wash twice with cell staining buffer (see step 4) then resuspend in 0.5 ml cell staining buffer. Analyze with flow cytometer using appropriate instrument settings.

NOTE: FOXP3 Fix/Perm buffer set is specifically developed and formulated for intracellular staining FOXP3 with minimum effect on surface fluorochrome staining and is highly recommended for optimal result of FOXP3 intracellular immunofluorescence staining.
Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent intracellular staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.
Application References:

1. Roncador G, et al. 2005 Eur. J. Immunol. 35:1681.
2. Yang ZZ, et al. 2006. Blood 107:3639.
3. Liu W, et al. 2006. J. Exp. Med. 203:1701.PubMed
4. Bollyky PL, et al. 2007. J. Immunol. 179:744.
5. Bell MP, et al. 2007. J. Immunol. 179:1893.
7. Tran DQ, et al. 2007. Blood doi:10.1182/blood-2007-06-094656. PubMed
8. Gao Q,et al.2007.J Clin Oncol.25:2586.PubMed
9. Pillai V,et al. 2008. Blood 111:463.PubMed
10. Zheng Y, et al. 2008. J. Immunol. 181:1683. PubMed
11. Zonios DI, et al. 2008.Blood112:287. PubMed
12. Kavanagh B, et al. 2008. BloodPubMed
13. Nevala WK, et al. 2009. Clin Cancer Res. 15:1931. PubMed
14. Grant J, et al. 2009. Cytometry B Clin Cytom. 76:69. PubMed
15. Nigam P, et al. 2010. J. Immunol. 184:1690. PubMed
16. Kmieciak M, et al. 2009. J. Transl. Med. 7:89. (ICFC) PubMed
17. Hartigan-O'Connor DJ,et al.2007.J Exp Med.204:2679. PubMed
18. Raghaven S, et al. 2009. Ann Rheum Dis. 68:1908. PubMed
Human peripheral blood lymphocytes were

Human peripheral blood lymphocytes were surface stained with CD4 PerCP and then intracellularly stained with 206D Alexa Fluor® 647 by using Alexa Fluor® 647 anti-human FOXP3 Flow Kit. Quadrant markers were set based on the staining of mouse IgG1, κ Alexa Fluor® 647 isotype control.



Description:

FOXP3 is a 50-55 kD transcription factor, also known as Forkhead box protein P3, Scurfin, JM2, or IPEX. It is proposed to be a master regulatory gene and more specific marker of T regulatory cells than most cell surface markers (such as CD4 and CD25). Transduced expression of FOXP3 in CD4+/CD25- cells has been shown to induce GITR, CD103, and CTLA4 and impart a T regulatory cell phenotype. FOXP3 is mutated in X-linked autoimmunity-allergic dysregulation syndrome (XLAAD or IPEX) in humans and in "scurfy" mice. Overexpression of FOXP3 has been shown to lead to a hypoactive immune state suggesting that this transcriptional factor is a central regulator of T cell activity. In human, unlike in mouse, two isoforms of FOXP3 have been reported: one (FOXP3) corresponding to the canonical full-length sequence; the other (FOXP3 δ2) lacking exon 2. The 206D antibody recognizes human FOXP3 epitope in the region of amino acids 105-235.
Other Names: Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
Structure: Forkhead/winged-helix transcription factor family, approximately 50 kD, contains zinc finger and forkhead domains
Distribution: Nuclear; expressed in T regulatory cells
Antigen References:

1. Hori S, et al. 2003. Science 299:1057.
2. Gandhi R, et al. 2010. Nat. Immunol. 11:846.

This product is provided under an agreement between Life Technologies Corporation and Sony Biotechnology Inc. and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications, and corresponding non-U.S. equivalents, owned by and/or licensed to Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research and/or development conducted by the buyer (whether the buyer is an academic or for profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to (1) use of the product or its components in manufacturing, (2) use of the product or its components for therapeutic, diagnostic or prophylactic purposes, or (3) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Molecular Probes Detection Technologies, Attn: Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  • 目录号：2200595
  • 品牌：Sony Biotechnology
  • 规格：25 tests
  • 目录价：￥6384.00
  • 市场价格：￥4149.60
  • 会员价格：￥3958.08
  • 数量：

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