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.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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™ Violet 750 is covered by one or more of the following US patents: 8,158,444; 8,802,450; 8,575,303; 8,455,613; 8,227,187; 8,841,072; 8,110,673.
Companion Products
The 25F.1D6 monoclonal antibody specifically recognizes CD366 which is also known as TIM-3 (T-cell immunoglobulin and mucin domain-containing 3, T-cell immunoglobulin mucin receptor 3, or T-cell membrane protein 3). CD366 (TIM-3) is an ~31 kDa type I transmembrane glycoprotein encoded by Havcr2 (Hepatitis A virus cellular receptor 2) that belongs to the TIM family within the Ig superfamily. CD366 (TIM-3) is comprised of one IgV-type domain followed by a serine/threonine-rich mucin stalk region in its extracellular region, a transmembrane segment, and a tyrosine phosphorylation motif in its cytoplasmic tail. CD366 (TIM-3) expression is upregulated on subpopulations of activated myeloid cells including macrophages, monocytes, dendritic cells (DC), microglia, mast cells as well as on Type-1 CD4+ (Th1-like) T cells, cytotoxic CD8+ T cells, regulatory T cells (Treg), and natural killer (NK) cells. CD366 (TIM-3) functions as an inhibitory receptor that helps maintain immunological homeostasis and self-tolerance. It may also serve an immune checkpoint molecule that inhibits antitumor immunity and promotes T cell exhaustion. Crosslinking of cell surface CD366 (TIM-3) by Galectin-9 binding downregulates Th1-like and CD8+ T cell responses and can promote Treg or myeloid-derived suppressor cells. CD366 (TIM-3) enables DC to bind phosphatidyl serine expressed by apoptotic cells, phagocytize these cells to suppress inflammation and promote antigen cross-presentation. CD366 (TIM-3) can also bind to high mobility group protein B1 (HMGB1) and inhibit stimulation of the immune response to nucleic acids released by dying tumor cells.
Development References (9)
-
Anderson AC, Joller N, Kuchroo VK. Lag-3, Tim-3, and TIGIT: Co-inhibitory Receptors with Specialized Functions in Immune Regulation.. Immunity. 2016; 44(5):989-1004. (Biology). View Reference
-
Chiba S, Baghdadi M, Akiba H, et al. Tumor-infiltrating DCs suppress nucleic acid-mediated innate immune responses through interactions between the receptor TIM-3 and the alarmin HMGB1.. Nat Immunol. 2012; 13(9):832-42. (Biology). View Reference
-
Monney L1, Sabatos CA, Gaglia JL, et al. Th1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease.. Nature. 2002; 415(6871):536-541. (Clone-specific: Flow cytometry). View Reference
-
Nakae S, Iikura M, Suto H, et al. TIM-1 and TIM-3 enhancement of Th2 cytokine production by mast cells.. Blood. 2007; 110(7):2565-8. (Biology). View Reference
-
Nakayama M, Akiba H, Takeda K, et al. Tim-3 mediates phagocytosis of apoptotic cells and cross-presentation. Blood. 2009; 113(16):3821-3830. (Biology). View Reference
-
Oikawa T1, Kamimura Y, Akiba H, et al. Preferential involvement of Tim-3 in the regulation of hepatic CD8+ T cells in murine acute graft-versus-host disease.. J Immunol. 2006; 177(7):4281-4287. (Biology). View Reference
-
Phong BL, Avery L, Sumpter TL, et al. Tim-3 enhances FcεRI-proximal signaling to modulate mast cell activation.. J Exp Med. 2015; 212(13):2289-304. (Biology). View Reference
-
Veenstra RG, Taylor PA, Zhou Q, et al. Contrasting acute graft-versus-host disease effects of Tim-3/galectin-9 pathway blockade dependent upon the presence of donor regulatory T cells.. Blood. 2012; 120(3):682-90. (Biology). View Reference
-
Zhu C, Anderson AC, Schubart A, et al. The Tim-3 ligand galectin-9 negatively regulates T helper type 1 immunity.. Nat Immunol. 2005; 6(12):1245-52. (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.