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 cell 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The RMT3-23 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 ~31 kDa type I transmembrane glycoprotein encoded by Havcr2 (Hepatitis A virus cellular receptor 2) that belongs to the TIM family within the immunoglobulin superfamily (IgSF). CD366 (TIM-3) is comprised of one Ig-like V-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. RMT3-23 reportedly recognizes CD366 (TIM-3) derived from either BALB/c or C57BL/6 mice that have different Havcr2 polymorphisms.
Development References (9)
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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
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Anderson AC, Xiao S, Kuchroo VK. Tim protein structures reveal a unique face for ligand binding.. Immunity. 2007; 26(3):273-5. (Biology). View Reference
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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
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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. (Clone-specific: Flow cytometry). View Reference
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Nakayama M, Akiba H, Takeda K, et al. Tim-3 mediates phagocytosis of apoptotic cells and cross-presentation. Blood. 2009; 113(16):3821-3830. (Clone-specific). View Reference
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Ngiow SF, von Scheidt B, Akiba H, Yagita H, Teng MW, Smyth MJ. Anti-TIM3 antibody promotes T cell IFN-γ-mediated antitumor immunity and suppresses established tumors.. Cancer Res. 2011; 71(10):3540-51. (Clone-specific: Flow cytometry). View Reference
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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. (Immunogen: Flow cytometry, Immunohistochemistry, In vivo exacerbation). View Reference
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Sakuishi K, Ngiow SF, Sullivan JM, et al. TIM3+FOXP3+ regulatory T cells are tissue-specific promoters of T-cell dysfunction in cancer.. Oncoimmunology. 2013; 2(4):e23849. (Clone-specific: Flow cytometry). View Reference
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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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.