Two-color flow cytometric analysis of human CCR10 expression on human peripheral blood lymphocytes. Whole blood was treated with BD PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were then stained with FITC Mouse Anti-Human CD3 antibody (Cat No. 555332/561806/561807) and either PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control (Cat. No. 550927; Left Panel) or PerCP-Cy™5.5 Mouse Anti-Human CCR10 antibody (Cat. No. 564772; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CCR10 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light- scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of human CCR10 expression on human peripheral blood lymphocytes. Whole blood was treated with BD PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were then stained with FITC Mouse Anti-Human CD3 antibody (Cat No. 555332/561806/561807) and either PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control (Cat. No. 550927; Left Panel) or PerCP-Cy™5.5 Mouse Anti-Human CCR10 antibody (Cat. No. 564772; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CCR10 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light- scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
製品詳細
BD Pharmingen™
C-C CKR-10, C-C CKR-10, G-protein coupled receptor 2, CCR-10, GPR2
Human (QC Testing)
Mouse C57BL/6 IgG2a, κ
Human CCR10 Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2738944
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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- 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.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
関連製品
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Stain Buffer (BSA) RUO
サイズ
500 mL
カタログ番号
554657
Lysing Solution 10X Concentrate RUO (GMP)
サイズ
100 mL
カタログ番号
349202
Lysing Buffer RUO
サイズ
100 mL
カタログ番号
555899
PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control RUO
サイズ
0.1 mg
カタログ番号
550927
FITC Mouse Anti-Human CD3 RUO
サイズ
100 Tests
カタログ番号
555332
564772 Rev. 2
抗体の詳細
1B5
The 1B5 monoclonal antibody specifically binds to CCR10 (CC-chemokine receptor 10). CCR10 is a seven transmembrane domain G-protein-coupled receptor that is also known as GPR-2 (G-protein coupled receptor 2). CCR10 is expressed by cells that mediate epithelial immunity including a small subset of T memory cells, IgA-secreting B cells, dermal endothelial cells and fibroblasts. It is also expressed on EBV-immortalized B cells. Chemokines CCL27 and CCL28 are the ligands for CCR10. CCR10 reportedly not only functions in regulating the homing and function of cells that mediate epithelial immunity. It may also regulate the epithelial localization, survival, and proliferation of tumor cells and facilitate their evasion from immune surveillance.
564772 Rev. 2
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
564772 Rev.2
引用＆参考文献
Development References (6)
-
Homey B, Wang W, Soto H, et al. Cutting edge: the orphan chemokine receptor G protein-coupled receptor-2 (GPR-2, CCR10) binds the skin-associated chemokine CCL27 (CTACK/ALP/ILC). J Immunol. 2000; 164(7):3465-3470. (Biology). View Reference
-
Jaimes MC, Rojas OL, Kunkel EJ, et al. Maturation and trafficking markers on rotavirus-specific B cells during acute infection and convalescence in children. J Virol. 2004; 78(20):10967-10976. (Clone-specific). View Reference
-
Marchese A, Docherty JM, Nguyen T, et al. Cloning of human genes encoding novel G protein-coupled receptors. Genomics. 1994; 23(3):609-618. (Biology). View Reference
-
Pan J, Kunkel EJ, Gosslar U, et al. A novel chemokine ligand for CCR10 and CCR3 expressed by epithelial cells in mucosal tissues. J Immunol. 2000; 165(6):2943-2949. (Biology). View Reference
-
Soler D, Humphreys TL, Spinola SM, Campbell JJ. CCR4 versus CCR10 in human cutaneous TH lymphocyte trafficking. Blood. 2003; 101(5):1677-1682. (Immunogen: Flow cytometry). View Reference
-
Xiong N, Fu Y, Hu S, Xia M, Yang J. CCR10 and its ligands in regulation of epithelial immunity and diseases. Protein Sci. 2012; 3(8):571-580. (Biology). View Reference
564772 Rev. 2
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.
