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 UIC2 monoclonal antibody specifically binds to an extracellular epitope of CD243, which is also known as ATP-binding cassette subfamily B member 1 (ABCB1), Multidrug resistance protein 1 (MDR1), or P-glycoprotein 1. CD243 is a transmembrane glycoprotein that spans the membrane 12 times. CD243 acts as an ATP-dependent efflux pump for a large variety of lipophilic molecules and drugs. This efflux activity has been suggested to lead to resistance to the drugs used in chemotherapy. CD243 is expressed by epithelial and endothelial cells, and at low levels by T cells, B cells, NK cells, and hematopoietic stem cells. It may be expressed at high levels by multidrug resistant (MDR) tumor cells. The UIC2 antibody reportedly inhibited the efflux of CD243 substrates from MDR cells and increased the cytotoxicity of certain CD243-transported drugs. The UIC2 antibody reportedly does not crossreact with mouse CD243.
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 (5)
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Mechetner EB1, Roninson IB. Efficient inhibition of P-glycoprotein-mediated multidrug resistance with a monoclonal antibody.. Proc Natl Acad Sci U S A. 1992; 89(13):5824-5828. View Reference
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Nagy H, Goda K, Arceci R, Cianfriglia M, Mechetner E, Szabó G. P-Glycoprotein conformational changes detected by antibody competition.. Eur J Biochem. 2001; 268(8):2416-20. View Reference
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Ritchie TK, Kwon H, Atkins WM. Conformational analysis of human ATP-binding cassette transporter ABCB1 in lipid nanodiscs and inhibition by the antibodies MRK16 and UIC2.. J Biol Chem. 2011; 286(45):39489-96. View Reference
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Szalóki G, Krasznai ZT, Tóth Á, et al. The strong in vivo anti-tumor effect of the UIC2 monoclonal antibody is the combined result of Pgp inhibition and antibody dependent cell-mediated cytotoxicity.. PLoS ONE. 2014; 9(9):e107875. View Reference
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Zhou Y, Gottesman MM, Pastan I. The extracellular loop between TM5 and TM6 of P-glycoprotein is required for reactivity with monoclonal antibody UIC2.. Arch Biochem Biophys. 1999; 367(1):74-80. 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.