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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 AA4.1 monoclonal antibody specifically recognizes the Early B Lineage antigen which is also known as CD93, AA4 antigen, Ly-68, and Complement component C1q receptor (C1qRp). This 130-140-kDa type I transmembrane glycoprotein is expressed on immature B lymphocytes in the adult bone marrow and on hematopoietic progenitors and stem cells in adult bone marrow, fetal liver, and embryonic yolk sac. Although CD93+ cells are most plentiful in adult mouse bone marrow, a smaller number of CD93+ cells which express lower CD93 levels can be detected in the adult spleen using bright fluorescent conjugates of the AA4.1 antibody or an amplified indirect immunofluorescent staining procedure. It has been observed that the staining pattern of the 493 monoclonal antibody is similar to that of the AA4.1 antibody, in that both antibodies precipitate molecules of the same molecular weight. Staining with the AA4.1 antibody is not blocked by the 493 antibody. These results suggest that the antibodies recognize separate epitopes on the same Early B Lineage antigen.
Development References (9)
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Allman D, Li J, Hardy RR. Commitment to the B lymphoid lineage occurs before DH-JH recombination. J Exp Med. 1999; 189(4):735-740. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Allman D, Lindsley RC, DeMuth W, Rudd K, Shinton SA, Hardy RR. Resolution of three nonproliferative immature splenic B cell subsets reveals multiple selection points during peripheral B cell maturation. J Immunol. 2001; 167(12):6834-6840. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Auerbach R, Huang H, Lu L. Hematopoietic stem cells in the mouse embryonic yolk sac. Stem Cells. 1996; 14(3):269-280. (Clone-specific). View Reference
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Jordan CT, McKearn JP, Lemischka IR. Cellular and developmental properties of fetal hematopoietic stem cells. Cell. 1990; 61(6):953-963. (Clone-specific: Flow cytometry). View Reference
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Lacaud G, Carlsson L, Keller G. Identification of a fetal hematopoietic precursor with B cell, T cell, and macrophage potential. Immunity. 1998; 9(6):827-838. (Clone-specific). View Reference
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Li YS, Wasserman R, Hayakawa K, Hardy RR. Identification of the earliest B lineage stage in mouse bone marrow. Immunity. 1996; 5(6):527-535. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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McKearn JP, Baum C, Davie JM. Cell surface antigens expressed by subsets of pre-B cells and B cells. J Immunol. 1984; 132(1):332-339. (Immunogen: Cell separation, Depletion, Flow cytometry). View Reference
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Paige CJ, Gisler RH, McKearn JP, Iscove NN. Differentiation of murine B cell precursors in agar culture. Frequency, surface marker analysis and requirements for growth of clonable pre-B cells. Eur J Immunol. 1984; 14(11):979-987. (Clone-specific). View Reference
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Szilvassy SJ, Cory S. Phenotypic and functional characterization of competitive long-term repopulating hematopoietic stem cells enriched from 5-fluorouracil-treated murine marrow. Blood. 1993; 81(9):2310-2320. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). 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.