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 1F2 mAb reacts with Crry/p65, a cell-surface protein that blocks complement activation and C3 deposition by both the classical and alternative pathways. Based on nucleotide sequence identity, Crry/p65 is the genetic homologue of human CR1 (Complement Receptor 1). However, it has been proposed that Crry/p65 is the functional homologue of human MCP (Membrane Cofactor Protein) and/or DAF (Decay-Accelerating Factor), due to the similar functions these proteins share and the fact that Crry/p65 does not display CR1's complement-receptor activity. Crry/p65 is expressed in most organs, such as thymus, kidney, uterus, skin, lung, pancreas, heart, stomach, large and small intestine, and spleen. Platelets, brain, and muscle express low, but detectable levels of Crry/p65. In addition, Crry/p65 is expressed in J774 cells, bone marrow-derived macrophages, and thioglycollate-elicited peritoneal macrophages, as determined by RT-PCR. Crry is crucial in the regulation of complement activation during early mouse embryonic development and essential in fetomaternal tolerance. The 1F2 mAb partially blocks Crry/p65-mediated complement protection in vitro.
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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Foley S, Li B, Dehoff M, Molina H, Holers VM. Mouse Crry/p65 is a regulator of the alternative pathway of complement activation. Eur J Immunol. 1993; 23(6):1381-1384. (Biology). View Reference
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Holers VM, Kinoshita T, Molina H. The evolution of mouse and human complement C3-binding proteins: divergence of form but conservation of function. Immunol Today. 1992; 13(6):231-236. (Biology). View Reference
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Kurtz CB, O'Toole E, Christensen SM, Weis JH. The murine complement receptor gene family. IV. Alternative splicing of Cr2 gene transcripts predicts two distinct gene products that share homologous domains with both human CR2 and CR1. J Immunol. 1990; 144(9):3581-3591. (Biology). View Reference
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Li B, Sallee C, Dehoff M, Foley S, Molina H, Holers VM. Mouse Crry/p65. Characterization of monoclonal antibodies and the tissue distribution of a functional homologue of human MCP and DAF. J Immunol. 1993; 151(8):4295-4305. (Immunogen). View Reference
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Martin BK, Weis JH. Murine macrophages lack expression of the Cr2-145 (CR2) and Cr2-190 (CR1) gene products. Eur J Immunol. 1993; 23(11):3037-3042. (Biology). View Reference
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Molina H, Wong W, Kinoshita T, Brenner C, Foley S, Holers VM. Distinct receptor and regulatory properties of recombinant mouse complement receptor 1 (CR1) and Crry, the two genetic homologues of human CR1. J Immunol. 1993; 175(1):121-129. (Biology). View Reference
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Xu C, Mao D, Holers VM, Palanca B, Cheng AM, Molina H. A critical role for murine complement regulator crry in fetomaternal tolerance. Science. 2000; 287(5452):498-501. (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.
