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
- 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.
- 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.
- 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.
- 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.
- 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 http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The RMT2-26 monoclonal antibody specifically recognizes T cell immunoglobulin and mucin domain-2 (TIM-2) which is also known as T-cell membrane protein 2, TIMD-2, or Tim2. TIM-2 is an ~55 kDa type I transmembrane glycoprotein that is encoded by Timd2 (T cell immunoglobulin and mucin domain containing 2) which belongs to the immunoglobulin gene superfamily. TIM-2 contains extracellular IgV and mucin domains and a cytoplasmic region with a conserved tyrosine phosphorylation motif which is involved in transmembrane signaling. A human ortholog for mouse TIM-2 has not been identified. TIM-2 is reportedly expressed on B cells with higher levels on activated B cells and germinal center B cells, type-2 T helper (Th2) cells, oligodendrocytes, and epithelial cells in the kidney and liver. It may serve as a receptor for Semaphorin 4A (Sema4A) or the heavy chain of ferritin (H-ferritin). The RMT2-26 antibody can reportedly block the binding of H-ferritin to TIM-2.
Development References (8)
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Abe Y, Kamachi F, Kawamoto T, et al. TIM-4 has dual function in the induction and effector phases of murine arthritis.. J Immunol. 2013; 191(9):4562-72. (Clone-specific: Flow cytometry). View Reference
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Chakravarti S, Sabatos CA, Xiao S, et al. Tim-2 regulates T helper type 2 responses and autoimmunity.. J Exp Med. 2005; 202(3):437-44. (Biology). View Reference
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Chen TT, Li L, Chung DH, et al. TIM-2 is expressed on B cells and in liver and kidney and is a receptor for H-ferritin endocytosis.. J Exp Med. 2005; 202(7):955-65. (Biology). View Reference
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Han J, Seaman WE, Di X, et al. Iron uptake mediated by binding of H-ferritin to the TIM-2 receptor in mouse cells.. PLoS ONE. 2011; 6(8):e23800. (Biology). View Reference
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Kawamoto T, Abe Y, Ito J, et al. Anti-T cell immunoglobulin and mucin domain-2 monoclonal antibody exacerbates collagen-induced arthritis by stimulating B cells.. Arthritis Res Ther. 2011; 13(2):R47. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Western blot). View Reference
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Kumanogoh A, Marukawa S, Suzuki K, et al.. Class IV semaphorin Sema4A enhances T-cell activation and interacts with Tim-2. Nature. 2002; 419(6907):629-633. (Biology). View Reference
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Rennert PD, Ichimura T, Sizing ID, et al. T cell, Ig domain, mucin domain-2 gene-deficient mice reveal a novel mechanism for the regulation of Th2 immune responses and airway inflammation.. J Immunol. 2006; 177(7):4311-21. (Biology). View Reference
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Rodriguez-Manzanet R, DeKruyff R, Kuchroo VK, Umetsu DT. The costimulatory role of TIM molecules. Immunol Rev. 2009; 229(1):259-270. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
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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.