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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 R1-2 monoclonal antibody specifically binds to α4 chain (CD49d), which is expressed as a heterdimer with either of two β, β1 or β7 (also known as βp). The α4β1 integrin (VLA-4, CD49d/CD29) is expressed on most peripheral lymphocytes, thymocytes, and monocytes; while the α4β7 integrin (LPAM-1) is expressed on peripheral lymphocytes, but on only a small subset of thymocytes. These integrins mediate a variety of cell-cell and cell-matrix interactions, recognizing the ligands VCAM-1 (CD106) and fibronectin. There is evidence that levels of VLA-4 expression regulate the transendothelial migration of T lymphocytes into inflamed tissues. Integrin α4β7 also preferentially binds to the mucosal vascular addressin, MAdCAM-1. The R1-2 antibody blocks some α4 integrin-mediated binding functions. In combination with mAb 9C10 (MFR4.B) (Cat. No. 553313), binding of VLA-4 expressing cells to VCAM-1 can be almost completely inhibited.
Development References (8)
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Andrew DP, Berlin C, Honda S, et al. Distinct but overlapping epitopes are involved in alpha 4 beta 7-mediated adhesion to vascular cell adhesion molecule-1, mucosal addressin-1, fibronectin, and lymphocyte aggregation. J Immunol. 1994; 153(9):3847-3861. (Biology). View Reference
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Berlin C, Berg EL, Briskin MJ, et al. Alpha 4 beta 7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell. 1993; 74(1):185-195. (Biology). View Reference
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Ferguson TA, Kupper TS. Antigen-independent processes in antigen-specific immunity. A role for alpha 4 integrin. J Immunol. 1993; 150(4):1172-1182. (Biology). View Reference
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Holzmann B, McIntyre BW, Weissman IL. Identification of a murine Peyer's patch--specific lymphocyte homing receptor as an integrin molecule with an alpha chain homologous to human VLA-4 alpha. Cell. 1989; 56(1):37-46. (Immunogen). View Reference
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Holzmann B, Weissman IL. Peyer's patch-specific lymphocyte homing receptors consist of a VLA-4-like alpha chain associated with either of two integrin beta chains, one of which is novel. EMBO J. 1989; 8(6):1735-1741. (Clone-specific: Immunoprecipitation, Inhibition). View Reference
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Kilshaw PJ, Murant SJ. Expression and regulation of beta 7(beta p) integrins on mouse lymphocytes: relevance to the mucosal immune system. Eur J Immunol. 1991; 21(10):2591-2597. (Biology). View Reference
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Kinashi T, Springer TA. Adhesion molecules in hematopoietic cells. Blood Cells. 1994; 20(1):25-44. (Biology). View Reference
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Romanic AM, Graesser D, Baron JL, Visintin I, Janeway CA Jr, Madri JA. T cell adhesion to endothelial cells and extracellular matrix is modulated upon transendothelial cell migration. Lab Invest. 1997; 76(1):11-23. (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.