Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
The P-3E10 monoclonal antibody specifically recognizes CD298, the β3 subunit of the Na+/K+ ATPase (Sodium/potassium-transporting ATPase subunit beta-3). CD298 is a ~45-55 kDa single-pass, type II membrane protein that is encoded by ATP1B3 (ATPase Na+/K+ transporting subunit beta 3) which belongs to the P-type ATPases superfamily. CD298 is widely expressed on lymphocytes, monocytes, granulocytes, platelets, and on other hematopoietic and nonhematopoietic cells and cell lines. The Na+/K+ ATPase is an integral membrane protein complex with enzymatic activity that mediates the active transport and exchange of sodium and potassium ions across the plasma membrane. This complex is composed of α and β subunits. The α subunit is a 10-membrane-spanning, catalytic protein that contains binding sites for Na +, K + and ATP. The α subunits are associated with the smaller, regulatory glycoprotein β subunits. Upon ATP hydrolysis, the Na+/K+ ATPase transports Na+ ions out of the cell in exchange for K+ ions that are transported in. This establishes a transmembrane electrochemical gradient that is essential for osmoregulation and for the transport of various nutrients and other molecules by cells. The P-3E10 antibody can reportedly inhibit the activation and proliferation of T cells and B cells.
Development References (5)
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Bouwer AL, Saunderson SC, Caldwell FJ, et al. NK cells are required for dendritic cell-based immunotherapy at the time of tumor challenge.. J Immunol. 2014; 192(5):2514-21. (Clone-specific: Flow cytometry). View Reference
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P. A. Gorer, S. Lyman, G. D. Snell. Studies on the Genetic and Antigenic Basis of Tumour Transplantation. Linkage between a Histocompatibility Gene and 'Fused' in Mice. Proc R Soc Lond B Biol Sci. 1948; 135(881):499-505. (Biology).
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Springer TA. Cell-surface differentiation in the mouse. Characterization of "jumping" and "lineage" antigens using xenogeneic rat monoclonal antibodies. In: Kennett RH, McKearn TJ, Bechtol KB, ed. Monoclonal antibodies. Hybridomas: A new dimension in biological analyses. New York and London: Plenum Press; 1980:185-217.
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Stallcup KC, Springer TA, Mescher MF. Characterization of an anti-H-2 monoclonal antibody and its use in large-scale antigen purification.. J Immunol. 1981; 127(3):923-30. (Immunogen: Flow cytometry, Immunoaffinity chromatography, Immunoprecipitation, Radioimmunoassay). View Reference
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Watanabe Y, Kuribayashi K, Miyatake S, et al. Exogenous expression of mouse interferon gamma cDNA in mouse neuroblastoma C1300 cells results in reduced tumorigenicity by augmented anti-tumor immunity.. Proc Natl Acad Sci USA. 1989; 86(23):9456-60. (Clone-specific: Flow cytometry). 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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.