Flow cytometric analysis of mouse IL-17F expression in resting or stimulated mouse EL4 thymoma cells or in polarized mouse Th17 cells. Mouse EL4 thymoma cells were either unstimulated or stimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) and Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 5 hours. Th17 cells were generated from Th17-polarizing mouse spleen cell cultures and were stimulated with PMA and Ionomycin with BD GolgiStop™ Protein Transport Inhibitor. The EL4 and Th17 cells were fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The unstimulated (Left Panel) and stimulated (Middle Panel) EL4 cells were stained with PerCP-Cy™5.5 Mouse Anti-Mouse IL-17F (Cat. No. 562194). To validate O79-289 antibody use for staining IL-17F in Th17 cells generated from primary mouse T cells, the fixed and permeabilized Th17-polarized cells were stained with purified O79-289 Mouse Anti-Mouse IL-17F antibody followed by PE Goat Anti-Mouse Ig (Cat. No. 550589) and with Alexa Fluor® 488 Rat Anti-Mouse IL-17A antibody (Cat. No. 560220) (Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of IL-17F versus cellular autofluorescence (measured in the FL1 channel) for EL4 cells or versus IL-17A for Th17 cells were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.
Flow cytometric analysis of mouse IL-17F expression in resting or stimulated mouse EL4 thymoma cells or in polarized mouse Th17 cells. Mouse EL4 thymoma cells were either unstimulated or stimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) and Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 5 hours. Th17 cells were generated from Th17-polarizing mouse spleen cell cultures and were stimulated with PMA and Ionomycin with BD GolgiStop™ Protein Transport Inhibitor. The EL4 and Th17 cells were fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The unstimulated (Left Panel) and stimulated (Middle Panel) EL4 cells were stained with PerCP-Cy™5.5 Mouse Anti-Mouse IL-17F (Cat. No. 562194). To validate O79-289 antibody use for staining IL-17F in Th17 cells generated from primary mouse T cells, the fixed and permeabilized Th17-polarized cells were stained with purified O79-289 Mouse Anti-Mouse IL-17F antibody followed by PE Goat Anti-Mouse Ig (Cat. No. 550589) and with Alexa Fluor® 488 Rat Anti-Mouse IL-17A antibody (Cat. No. 560220) (Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of IL-17F versus cellular autofluorescence (measured in the FL1 channel) for EL4 cells or versus IL-17A for Th17 cells were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.
製品詳細
BD Pharmingen™
Il17f; interleukin 17F; Interleukin-17F
Mouse (QC Testing)
Mouse BALB/c IgG1, κ
Mouse IL-17F Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_11153316
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
関連製品
Fixation/Permeabilization Kit RUO
サイズ
250 Tests
カタログ番号
554714
Protein Transport Inhibitor (Containing Monensin) RUO
サイズ
0.7 mL
カタログ番号
554724
PE Goat Anti-Mouse Ig (Multiple Adsorption) RUO
サイズ
0.2 mg
カタログ番号
550589
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Alexa Fluor® 488 Rat anti-Mouse IL-17A RUO
サイズ
0.1 mg
カタログ番号
560220
562194 Rev. 1
抗体の詳細
O79-289
The O79-289 monoclonal antibody specifically binds to the inflammatory cytokine protein, Interleukin-17 (IL-17F). IL-17F is a member of the IL-17 family of cytokines. Among IL-17 family members, IL-17F has the highest amino acid sequence homology to IL-17A. IL-17F is produced by activated CD4+ T helper (Th17) cells, CD8+ T (Tc17) cells and γδ T cells. IL-17F can be secreted as homodimers or as heterodimers with IL-17A. IL-17F and IL-17A have overlapping functions such as inducing epithelial cells and fibroblasts to produce proinflammatory cytokines and chemokines including IL-6, GM-CSF, CXCL1, CCL2, and CCL7. These factors attract and activate neutrophils and other cell types that mediate protective responses against pathogenic microbes or pathologic allergic or autoimmune diseases. IL-17 gene knockout studies have shown that IL-17F and IL-17A have independent functions as well. IL-17F and IL-17A exert their biological function by binding to and signaling through IL-17 receptors comprised of the transmembrane receptor subunits, IL-17RA (CD217) and IL-17RC.
562194 Rev. 1
フォーマットの詳細
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
562194 Rev.1
引用＆参考文献
Development References (8)
-
Chang SH, Dong C. IL-17F: Regulation, signaling and function in inflammation. Cytokine. 2009; 46(1):7-11. (Biology). View Reference
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Dong C. Th17 cells: Current understanding of their generation and regulation. Eur J Immunol. 2009; 39(3):640-644. (Biology). View Reference
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Hamada H, Garcia-Hernandez MdlL, Reome JB, et al. Tc17, a unique subset of CD8 T cells that can protect against lethal influenza challenge. J Immunol. 2009; 182(6):3469-3481. (Biology). View Reference
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Ishigame H, Kakuta S, Nagai T, et al. Differential roles of interleukin-17A and -17F in host defense against mucoepithelial bacterial infection and allergic responses. Immunity. 2009; 30(1):108-119. (Biology). View Reference
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Martinez GJ, Zhang Z, Chung Y, et al. Smad3 differentially regulates the induction of regulatory and inflammatory T cell differentiation. J Biol Chem. 2009; 284(51):35283-35286. (Biology). View Reference
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Oda N, Canelos PB, Essayan DM, Plunkett BA, Myers AC, Huang SK. Interleukin-17F induces pulmonary neutrophilia and amplifies antigen-induced allergic response. Am J Respir Crit Care Med. 2005; 171(1):12-13. (Biology). View Reference
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Sutton CE, Lalor SJ, Sweeney CM, Brereton CF, Lavelle EC, Mills KH. Interleukin-1 and IL-23 induce innate IL-17 production from gammadelta T cells, amplifying Th17 responses and autoimmunity. Immunity. 2009; 31(2):331-341. (Biology). View Reference
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Yang XO, Chang SH, Park H, et al. Regulation of inflammatory responses by IL-17F. J Exp Med. 2008; 205(5):1063-1075. (Biology). View Reference
562194 Rev. 1
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
