Two-color analysis of CD55 expression on splenic B lymphocytes. C57BL/6 splenocytes were pre-incubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 (Cat. No. 553141/553142), stained with FITC anti-mouse CD45R/B220 (Cat. No. 553087/553088) and either PE Hamster IgG3, λ isotype control (Cat. No. 553980, left panel) or PE Hamster anti-Mouse CD55 (Cat. No. 58037, right panel). Dead cells were excluded by staining with propidium iodide (Cat. No. 556463), and the total viable leukocytes are displayed. The data demonstrates that CD55 is expressed on the majority of splenocytes, with the greatest density on B lymphocytes. Flow cytometry was performed on a BD FACScalibur™ flow cytometry system.
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Two-color analysis of CD55 expression on splenic B lymphocytes. C57BL/6 splenocytes were pre-incubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 (Cat. No. 553141/553142), stained with FITC anti-mouse CD45R/B220 (Cat. No. 553087/553088) and either PE Hamster IgG3, λ isotype control (Cat. No. 553980, left panel) or PE Hamster anti-Mouse CD55 (Cat. No. 58037, right panel). Dead cells were excluded by staining with propidium iodide (Cat. No. 556463), and the total viable leukocytes are displayed. The data demonstrates that CD55 is expressed on the majority of splenocytes, with the greatest density on B lymphocytes. Flow cytometry was performed on a BD FACScalibur™ flow cytometry system.
Product Details
BD Pharmingen™
Cd55; DAF; decay-accelerating factor
Mouse (QC Testing)
Armenian Hamster IgG3, λ
GPI-DAF transfected CHO cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_397002
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Propidium Iodide Staining Solution RUO
Size
2 mL
Cat No.
556463
PE Hamster IgG3 λ1 Isotype Control RUO
Size
0.1 mg
Cat No.
553980
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558037 Rev. 2
Antibody Details
RIKO-5
The RIKO-5 monoclonal antibody specifically recognizes the membrane-associated complement regulator CD55, also known as decay-accelerating factor (DAF). CD55 is expressed on the surface of erythrocytes and most splenic leukocytes, including B and T lymphocytes and CD11b-positive splenocytes, but it is not detected on thymocytes. Immunohistochemical staining (using a different monoclonal antibody) demonstrates that CD55 is expressed on vascular endothelium and many other cells types in a wide variety of organs. In the mouse, there are two isoforms of CD55, transmembrane and GPI-anchored, that are recognized by the RIKO-5 antibody. Both isoforms are able to protect cells from complement attack by inhibiting the deposition of C3 on antibody-sensitized cells, but the RIKO-5 antibody is not able to neutralize this activity. CD97, a member of the EGF-TM7 family that is widely expressed in mouse tissues, is another ligand for CD55.
558037 Rev. 2
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
558037 Rev.2
Citations & References
Development References (7)
-
Ahmad SR, Lidington EA, Ohta R, et al. Decay-accelerating factor induction by tumour necrosis factor-alpha, through a phosphatidylinositol-3 kinase and protein kinase C-dependent pathway, protects murine vascular endothelial cells against complement deposition. Immunology. 2003, October; 110(2):258-268. (Biology). View Reference
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Leemans JC, te Velde AA, Florquin S, et al. The epidermal growth factor-seven transmembrane (EGF-TM7) receptor CD97 is required for neutrophil migration and host defense. J Immunol. 2004, January; 172(2):1125-1131. (Biology). View Reference
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Lin F, Fukuoka Y, Spicer A, et al. Tissue distribution of products of the mouse decay-accelerating factor (DAF) genes. Exploitation of a Daf1 knock-out mouse and site-specific monoclonal antibodies. Immunology. 2001, October; 104(2):215-225. (Biology). View Reference
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Miwa T, Maldonado MA, Zhou L, et al. (CD55) exacerbates autoimmune disease development in MRL/lpr mice. Am J Pathol. 2002, September ; 161(3):1077-1086. (Biology). View Reference
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Miwa, T., X. Sun, et al. Characterization of glycosylphosphatidylinositol-anchored decay accelerating factor (GPI-DAF) and transmembrane DAF gene expression in wild-type and GPI-DAF gene knockout mice using polyclonal and monoclonal antibodies with dual or single specificity.. Immunology. 2001; 104:207-214. (Biology). View Reference
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Ohta R, Imai M, Fukuoka Y, Miwa T, Okada N, Okada H. Characterization of mouse DAF on transfectant cells using monoclonal antibodies which recognize different epitopes. Microbiol Immunol. 1999; 43(11):1045-1056. (Biology). View Reference
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Yamada K, Miwa T, Liu J, Nangaku M, Song WC. Critical protection from renal ischemia reperfusion injury by CD55 and CD59. J Immunol. 2004, March; 172(6):3869-3875. (Biology: Flow cytometry). View Reference
558037 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.
