BD OptiBuild™ BUV615 Mouse Anti-Human MUC1 (CD227)
Clone HMFG2 (also known as HMFG-2, 3.14.A3 or F314AE11)
(RUO)
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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
Companion Products
The HMFG2 (Milk Fat Globule 2) monoclonal antibody, formerly known as clone 3.14.A3, specifically binds to Mucin-1 (MUC1) which is also known as CD227, Episialin, Breast carcinoma-associated antigen DF3, Cancer antigen 15-3, H23 antigen, Peanut reactive urinary protein (PUM), Polymorphic epithelial mucin (PEM), or Epithelial membrane antigen (EMA). MUC1 (CD227) is a heterodimeric cell surface glycoprotein that belongs to the epithelial mucin family. As a result of proteolytic cleavage, MUC1 (CD227) is comprised of a large, extracellular N-terminal alpha subunit that contains a domain of 20 amino-acid tandem repeats and functions in cellular adhesion. The smaller transmembrane C-terminal beta subunit contains a cytoplasmic domain that is involved in cell signaling. MUC1 (CD227) is variably expressed on the surfaces of normal and malignant glandular and ductal epithelial cells, myeloma cells, and some hematopoietic cell lineages including subsets of T cells, B cells, monocytes and dendritic cells. Soluble forms of MUC1 (CD227) may arise by shedding from the cell surface or by secretion of forms derived from alternative RNA splicing. MUC1 (CD227) plays roles in the provision of protective barrier function, the regulation of cellular adhesion and migration, and the transduction of multiple signal pathways. The HMFG2 antibody recognizes epitopes within the tandem repeat domain of MUC1 (CD227).
The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).
Development References (7)
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Arklie J, Taylor-Papadimitrious J, Bodmer W, Egan M, Millis R. Differentiation antigens expressed by epithelial cells in the lactating breast are also detectable in breast cancers.. Int J Cancer. 1981; 28(1):23-9. (Clone-specific: Immunohistochemistry). View Reference
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Burchell J, Durbin H, Taylor-Papadimitriou J. Complexity of expression of antigenic determinants, recognized by monoclonal antibodies HMFG-1 and HMFG-2, in normal and malignant human mammary epithelial cells.. J Immunol. 1983; 131(1):508-13. (Clone-specific: Radioimmunoassay, Western blot). View Reference
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Burchell J, Taylor-Papadimitriou J. Effect of modification of carbohydrate side chains on the reactivity of antibodies with core-protein epitopes of the MUC1 gene product.. Epithelial Cell Biol. 1993; 2(4):155-62. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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Correa I, Plunkett T, Vlad A, et al. Form and pattern of MUC1 expression on T cells activated in vivo or in vitro suggests a function in T-cell migration.. Immunology. 2003; 108(1):32-41. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
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McGuckin MA, MacDonald KP, Tran M, Wykes M, Hart DNJ. MUC1 Epithelial mucin: expression by normal hematopoietic cells. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:496-499.
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McGuckin MA. CD227 (MUC1) Summary and Workshop Report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:494-496.
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Taylor-Papadimitriou J, Peterson JA, Arklie J, Burchell J, Ceriani RL, Bodmer WF. Monoclonal antibodies to epithelium-specific components of the human milk fat globule membrane: production and reaction with cells in culture.. Int J Cancer. 1981; 28(1):17-21. (Immunogen: Immunoprecipitation, Radioimmunoassay). View Reference
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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.
