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 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 CompBead to ensure that BD Comp beads 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).
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).
- BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
Companion Products
The 1G8 monoclonal antibody specifically recognizes Endothelial cell-selective adhesion molecule (ESAM). ESAM is a ~55 kDa single-pass type I transmembrane glycoprotein that is encoded by ESAM (Endothelial cell-specific adhesion molecule) which belongs to the immunoglobulin supergene superfamily (IgSF). ESAM contains an IgV-type and IgC2-type domain in its extracellular region followed by a transmembrane sequence and cytoplasmic tail. ESAM is strongly expressed on platelets, megakaryocytes, and endothelial cells at interendothelial cell junctions as well as dendritic cells. Through homophilic interactions, this interendothelial cell adhesion molecule may play roles in regulating vascular permeability and in the extravasation of leucocytes, eg, neutrophils, through blood vessel walls. ESAM is also reportedly expressed on hematopoietic stem cells (HSCs). ESAM-positive cell populations are enriched for multipotent myeloid erythroid progenitors and primitive progenitors with lymphopoietic activity.
The antibody was conjugated to BD Horizon BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. With an Ex Max near 348 nm and an Em Max near 395 nm, BD Horizon BUV395 can be excited by the ultraviolet laser (355 nm) laser and detected with a 379/28 filter. This dye has been exclusively developed by BD Biosciences as an optimal dye for use on instruments equipped with the ultraviolet laser and has virtually no spillover into any other detector.
Development References (7)
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Duong CN, Nottebaum AF, Butz S, et al. Interference With ESAM (Endothelial Cell-Selective Adhesion Molecule) Plus Vascular Endothelial-Cadherin Causes Immediate Lethality and Lung-Specific Blood Coagulation. Arterioscler Thromb Vasc Biol. 2020; 40(2):378-393. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Western blot). View Reference
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Fujita K, Chakarov S, Kobayashi T, et al. Cell-autonomous FLT3L shedding via ADAM10 mediates conventional dendritic cell development in mouse spleen. Proc Natl Acad Sci U S A. 2019; 116(29):14714-14723. (Clone-specific: Flow cytometry). View Reference
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Hirata Ki, Ishida T, Penta K, et al. Cloning of an immunoglobulin family adhesion molecule selectively expressed by endothelial cells.. J Biol Chem. 2001; 276(19):16223-31. (Biology). View Reference
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Kirkling ME, Cytlak U, Lau CM, et al. Notch Signaling Facilitates In Vitro Generation of Cross-Presenting Classical Dendritic Cells. Cell Rep. 2018; 23(12):3658-3672. (Clone-specific: Flow cytometry). View Reference
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Lewis KL, Caton ML, Bogunovic M, et al. Notch2 receptor signaling controls functional differentiation of dendritic cells in the spleen and intestine. Immunity. 2011; 35(5):780-791. (Biology). View Reference
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Nasdala I, Wolburg-Buchholz K, Wolburg H, et al. A transmembrane tight junction protein selectively expressed on endothelial cells and platelets.. J Biol Chem. 2002; 277(18):16294-303. (Immunogen: Electron microscopy, ELISA, Flow cytometry, Fluorescence microscopy, Immunoaffinity chromatography, Immunofluorescence, Immunohistochemistry, Western blot). View Reference
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Sudo T, Yokota T, Oritani K, et al. The endothelial antigen ESAM monitors hematopoietic stem cell status between quiescence and self-renewal. J Immunol. 2012; 189(1):200-210. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). 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.