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 8A-1 monoclonal antibody specifically binds to CD148, which is also known as Protein tyrosine phosphatase receptor type J (Ptprj), Receptor-type tyrosine-protein phosphatase eta (R-PTP-eta, PTPη), or Density-enhanced phosphatase-1 (DEP-1). CD148 is a receptor-like tyrosine phosphatase that plays important roles in regulating cell growth, proliferation, differentiation, adhesion, and migration. It is expressed on a wide variety of cell types including epithelial cells, endothelial cells, and fibroblasts. For cells of hemopoietic origin, it is variably expressed on macrophages, neutrophils, dendritic cells, and on developing and mature B cells. CD148 is expressed on platelets and can regulate platelet activation. Although expressed at low levels on thymocytes and mature T cells, its expression is upregulated upon T cell receptor (TCR)-mediated stimulation and may play a role in regulating TCR signaling. CD148, along with CD45, are critical protein tyrosine phosphatases for regulating Src family kinase signaling networks in immune cells.
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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Cordoba SP, Choudhuri K, Zhang H, et al. The large ectodomains of CD45 and CD148 regulate their segregation from and inhibition of ligated T-cell receptor. Blood. 2013; 121(21):4295-4302. (Biology).
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Dave RK, Naylor AJ, Young SP et al. Differential expression of CD148 on leukocyte subsets in inflammatory arthritis. Arthritis Res Ther. 2013; 15(5):R108. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
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Katsumoto TR, Kudo M, Chen C et al. The phosphatase CD148 promotes airway hyperresponsiveness through SRC family kinases. J Clin Invest. 2013; 123(5):2037-2048. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
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Lin J, Zhu JW, Baker JE, Weiss A. Regulated expression of the receptor-like tyrosine phosphatase CD148 on hemopoietic cells. J Immunol. 2004; 173(4):2324-2330. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
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Senis YA, Tomlinson MG, Ellison S, Mazharian A et al. The tyrosine phosphatase CD148 is an essential positive regulator of platelet activation and thrombosis. Blood. 2009; 113(20):4942-4954. (Clone-specific: Flow cytometry, Western blot). 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.