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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
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The 346-11A monoclonal antibody specifically recognizes an epitope at the beginning of the cysteine-rich repeat region of the 200-kDa integrin β4 chain (CD104), which is found on the cell surface as a heterodimeric complex with the integrin α6 chain (CD49f). The α6β4 (CD49f/CD104) complex binds to laminins and is expressed on the basal surface of a variety of epithelial cell types, particularly on stratified squamous epithelia, and is also found in peripheral nerves, in certain subsets of endothelial cells, and on immature thymocytes. It has also been identified on a number of tumor tissues and participates in tumor progression events. Localization of both human and mouse epidermal α6β4 integrin to hemidesmosomes suggests that this heterodimer plays a role in epidermal adhesion to the basement membrane.
Development References (8)
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Giancotti FG. Signal transduction by the alpha 6 beta 4 integrin: charting the path between laminin binding and nuclear ev. J Cell Sci. 1996; 109(6):1165-1172. (Biology). View Reference
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Kennel SJ, Foote LJ, Falcioni R, et al. Analysis of the tumor-associated antigen TSP-180. Identity with alpha 6-beta 4 in the integrin superfamily. J Biol Chem. 1989; 264(26):15515-15521. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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Kennel SJ, Foote LJ, Flynn KM. Tumor antigen on benign adenomas and on murine lung carcinomas quantitated by a two-site monoclonal antibody assay. Cancer Res. 1986; 46(2):707-712. (Immunogen: Blocking, Immunoprecipitation, Radioimmunoassay). View Reference
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Morena A, Riccioni S, Marchetti A, et al. Expression of the beta 4 integrin subunit induces monocytic differentiation of 32D/v-Abl cells. Blood. 2002; 100(1):96-106. (Biology). View Reference
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Salmivirta K, Gullberg D, Hirsch E, Altruda F, Ekblom P. Integrin subunit expression associated with epithelial-mesenchymal interactions during murine tooth development. Dev Dyn. 1996; 205(2):104-113. (Clone-specific: Immunohistochemistry). View Reference
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Sonnenberg A, Calafat J, Janssen H, et al. Integrin alpha 6/beta 4 complex is located in hemidesmosomes, suggesting a major role in epidermal cell-basement membrane adhesion.. J Cell Biol. 1991; 113(4):907-17. (Clone-specific: Immunohistochemistry). View Reference
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Sonnenberg A, Linders CJ, Daams JH, Kennel SJ. The alpha 6 beta 1 (VLA-6) and alpha 6 beta 4 protein complexes: tissue distribution and biochemical properties. J Cell Sci. 1990; 96(2):207-217. (Clone-specific: Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
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van der Neut R, Krimpenfort P, Calafat J, Niessen CM, Sonnenberg A. Epithelial detachment due to absence of hemidesmosomes in integrin beta 4 null mice. Nat Genet. 1996; 13(3):366-369. (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.