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Flow cytometric analysis using BD OptiBuild™ RB780 Rat Anti-Mouse CD26 antibody (Cat. No. 755929; solid line histogram) on viable C57BL/6 mouse splenocytes, with corresponding Isotype Control (Cat. No. 568698; dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD OptiBuild™ RB780 Rat Anti-Mouse CD26
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The H194-112 monoclonal antibody specifically binds to CD26, which is also known as, Thymocyte-activating molecule (THAM), or dipeptidyl peptidase IV (DPP IV, Dpp4). CD26 is a ~220- kDa dimer formed of identical type-II transmembrane core polypeptides which undergo variable post-translational modifications. It is a multi-functional molecule with both ectopeptidase and signal-transducing activities. Studies with specific DPP IV inhibitors suggest that the enzymatic activity is involved in the mediation of T-cell activation events. The expression of CD26 is developmentally regulated in the thymus. Resting lymphoid cells of the bone marrow and peripheral B and T lymphocytes express low levels of CD26; bone-marrow and peritoneal myeloid cells do not. CD26 is also found on epithelial cells in the kidney, liver, small intestine, and lung. Cross-linked H194-112 mAb induces proliferation of immature and mature thymocytes in the presence of either IL-1 plus IL-2 or PMA; addition of IL-2 or IL-4 to PMA further enhances the activation.
Development References (7)
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Fleischer B. CD26: a surface protease involved in T-cell activation. Immunol Today. 1994; 15(4):180-184. (Biology). View Reference
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Gorvel JP, Vivier I, Naquet P, Brekelmans P, Rigal A, Pierres M. Characterization of the neutral aminopeptidase activity associated to the mouse thymocyte-activating molecule. J Immunol. 1990; 144(8):2899-2907. (Biology). View Reference
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Marguet D, Bernard AM, Vivier I, Darmoul D, Naquet P, Pierres M. cDNA cloning for mouse thymocyte-activating molecule. A multifunctional ecto-dipeptidyl peptidase IV (CD26) included in a subgroup of serine proteases. J Biol Chem. 1992; 267(4):2200-2208. (Biology). View Reference
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Naquet P, MacDonald HR, Brekelmans P. A novel T cell-activating molecule (THAM) highly expressed on CD4-CD8- murine thymocytes. J Immunol. 1988; 141(12):4101-4109. (Immunogen: Activation). View Reference
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Naquet P, Vivier I, Gorvel JP. Activation of mouse T lymphocytes by a monoclonal antibody to a developmentally regulated surface aminopeptidase (THAM). Immunol Rev. 1989; 111:177-193. (Clone-specific: Activation). View Reference
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Reinhold D, Bank U, Buhling F. Inhibitors of dipeptidyl peptidase IV (DP IV, CD26) induces secretion of transforming growth factor-beta 1 (TGF-beta 1) in stimulated mouse splenocytes and thymocytes. Immunol Lett. 1997; 58(1):29-35. (Biology: Activation). View Reference
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Vivier I, Marguet D, Naquet P . Evidence that thymocyte-activating molecule is mouse CD26 (dipeptidyl peptidase IV). J Immunol. 1988; 147(2):447-454. (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.