Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).
When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.
For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Product Notices
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
Companion Products
The R139 monoclonal antibody specifically binds to CD93 which is also known as Complement component C1q receptor (C1qR), C1q receptor 1 (C1qR1), or Matrix-remodeling-associated protein 4 (MXRA4). The immunogen used to generate the R139 hybridoma was a preparation of CD93 protein. Human CD93 is a transmembrane glycoprotein that is highly expressed on monocytes, macrophages, granulocytes, and endothelial cells but not on T and B lymphocytes. CD93 is also known as the C1q/MBL/SPA Receptor as it binds C1q, the recognition subunit of the first component (C1) of the complement pathway, as well as MBL (Mannose-binding-lectin) and SPA (Pulmonary Surfactant Protein A). Human C1qRp is involved in the C1q-mediated enhancement of phagocytosis. R139 is suitable to detect CD93 expression on cells of myeloid lineage by flow cytometry, and CD93 in cellular lysates by Western blotting or immunoprecipitation. In addition, R139 reportedly neutralizes C1q-mediated enhancement of phagocytosis. CD93 has also been reported to define a human stem cell population with hematopoietic and hepatic potential.
The antibody was conjugated to BD Horizon™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.
Development References (7)
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Danet GH, Luongo JL, Butler G, et al. C1qRp defines a new human stem cell population with hematopoietic and hepatic potential.. Proc Natl Acad Sci USA. 2002; 99(16):10441-5. (Biology). View Reference
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Guan E, Robinson SL, Goodman EB, Tenner AJ. Cell-surface protein identified on phagocytic cells modulates the C1q-mediated enhancement of phagocytosis. J Immunol. 1994; 152(8):4005-4016. (Immunogen: Bioassay, ELISA, Flow cytometry, Immunoprecipitation, Inhibition, Neutralization, Western blot). View Reference
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Guan EN, Burgess WH, Robinson SL, Goodman EB, McTigue KJ, Tenner AJ. Phagocytic cell molecules that bind the collagen-like region of C1q. Involvement in the C1q-mediated enhancement of phagocytosis. J Biol Chem. 1991; 266(30):20345-20355. (Immunogen: Functional assay, Immunoprecipitation, Inhibition, Western blot). View Reference
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Nepomuceno RR, Henschen-Edman AH, Burgess WH, Tenner AJ. cDNA cloning and primary structure analysis of C1qR(P), the human C1q/MBL/SPA receptor that mediates enhanced phagocytosis in vitro. Immunity. 1997; 6(2):119-129. (Clone-specific: Western blot). View Reference
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Nepomuceno RR, Ruiz S, Park M, Tenner AJ. C1qRP is a heavily O-glycosylated cell surface protein involved in the regulation of phagocytic activity. J Immunol. 1999; 162(6):3583-3589. (Clone-specific: Flow cytometry, Functional assay, Immunoprecipitation, Inhibition). View Reference
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Nepomuceno RR, Tenner AJ. C1qRP, the C1q receptor that enhances phagocytosis, is detected specifically in human cells of myeloid lineage, endothelial cells, and platelets. J Immunol. 1998; 160(4):1929-1935. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
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Tenner AJ. C1q receptors: regulating specific functions of phagocytic cells. Immunobiology. 1998; 199(2):250-264. (Biology). View Reference
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.