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Preparation and Storage
推奨アッセイ手順
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.
関連製品
The Cy34.1 monoclonal antibody specifically binds to the B-lymphocyte differentiation antigen CD22 on strains having the Lyb-8.2 alloantigen (e.g., A, BALB/c, CBA, C3H/He, C57BL, C57L, C58, SJL, SWR, but not AKR, DBA/1, DBA/2, NZB, PL). CD22 is expressed at high levels on mature peripheral B lymphocytes (follicular and marginal zone), B-1 cells (CD5+ B cells), and plasma cells. It is a member of the Ig gene superfamily and associates with the B-cell antigen receptor. Its sialic acid- binding immunoglobulin-like lectin (siglec) extracellular region mediates B-cell adhesion to ligands on endothelial cells in the bone marrow. Its intracellular domain is phosphorylated after cross-linking of antigen receptor or MHC class II antigen. It is involved in negative regulation of B-cell activation and protection from autoimmunity. B-cell proliferative responses to LPS or anti-mouse Ig µ chain are augmented in the presence of Cy34.1 mAb.
Development References (11)
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Doody GM, Justement LB, Delibrias CC. A role in B cell activation for CD22 and the protein tyrosine phosphatase SHP. Science. 1995; 269(5221):242-244. (Biology). View Reference
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Erickson LD, Tygrett LT, Bhatia SK, Grabstein KH, Waldschmidt TJ. Differential expression of CD22 (Lyb8) on murine B cells. Int Immunol. 1996; 8(7):1121-1129. (Biology). View Reference
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Law CL, Sidorenko SP, Clark EA. Regulation of lymphocyte activation by the cell-surface molecule CD22. Immunol Today. 1994; 15(9):442-449. (Biology). View Reference
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Law CL, Torres RM, Sundberg HA. Organization of the murine Cd22 locus. Mapping to chromosome 7 and characterization of two alleles. J Immunol. 1993; 151(1):175-187. (Clone-specific). View Reference
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Mary C, Laporte C, Parzy D. Dysregulated expression of the Cd22 gene as a result of a short interspersed nucleotide element insertion in Cd22a lupus-prone mice. J Immunol. 2000; 165(6):2987-2996. (Biology). View Reference
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Nitschke L, Floyd H, Ferguson DJ, Crocker PR. Identification of CD22 ligands on bone marrow sinusoidal endothelium implicated in CD22-dependent homing of recirculating B cells. J Exp Med. 1999; 189(9):1513-1518. (Biology). View Reference
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O'Keefe TL, Williams GT, Davies SL, Neuberger MS. Hyperresponsive B cells in CD22-deficient mice. Science. 1996; 274(5288):798-801. (Biology). View Reference
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Stall AM, Wells SM. FACS analysis of murine B-cell populations. In: Herzenberg LA, Weir DM, Blackwell C, ed. Weir's Handbook of Experimental Immunology. Blackwell Science Publishers; 1997:63.1-63.17.
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Stoddart A, Ray RJ, Paige CJ. Analysis of murine CD22 during B cell development: CD22 is expressed on B cell progenitors prior to IgM. Int Immunol. 1997; 9(10):1571-1579. (Biology). View Reference
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Symington FW, Subbarao B, Mosier DE, Sprent J. Lyb-8.2: A new B cell antigen defined and characterized with a monoclonal antibody. Immunogenetics. 1982; 16(5):381-391. (Immunogen: Immunoprecipitation). 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.
