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 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.
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.e6. (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):780-91. (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):200-10. (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.