Flow cytometric analysis using BD OptiBuild™ RY703 Rat Anti-Mouse CD43 antibody (Cat. No. 771748; solid line histogram) on viable BALB/c Mouse bone marrow cells, with Autofluorescence Control (dashed line histogram). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Flow cytometric analysis using BD OptiBuild™ RY703 Rat Anti-Mouse CD43 antibody (Cat. No. 771748; solid line histogram) on viable BALB/c Mouse bone marrow cells, with Autofluorescence Control (dashed line histogram). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Product Details
BD OptiBuild™
Spn; Sialophorin; Leukosialin; Ly-48; Ly48; Galgp; LEUK
Mouse (Tested in Development)
Rat DA x LOU IgG2a, κ
Mouse Plasmacytoma MOPC-315
Flow cytometry (Qualified)
0.2 mg/ml
20737
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
RY703 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571454
771748 Rev. 1
Antibody Details
S7
The S7 monoclonal antibody specifically binds to the 115 kDa glycosylated form of CD43 (Ly-48, Leukosialin). CD43 is expressed on IL-7-responsive pro-B cells, plasma cells, peritoneal and splenic CD5+ B cells (B-1 cells), granulocytes, monocytes, macrophages, platelets, natural killer cells, thymocytes, peripheral T cytotoxic/suppressor cells, and most T helper cells, but not resting conventional peripheral B cells. CD43 expression has also been detected on pluripotent hematopoietic stem cells and myeloid, lymphoid, and NK-cell progenitors in the bone marrow. Studies of CD43-deficient mice indicate that CD43 participates in the negative regulation of T-cell activation and adhesion.
771748 Rev. 1
Format Details
RY703
The BD Horizon RealYellow™ 703 (RY703) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 703-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY703 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY703 can be used as an alternative to PE-Cy5.5 and we recommend using an optical filter centered near 700-nm (eg, a 695/40-nm bandpass filter).
RY703
Yellow-Green 561 nm
557 nm
703 nm
771748 Rev.1
Citations & References
View product citations for antibody "771748" on CiteAb
Development References (9)
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Gulley ML, Ogata LC, Thorson JA, Dailey MO, Kemp JD. Identification of a murine pan-T cell antigen which is also expressed during the terminal phases of B cell differentiation. J Immunol. 1988; 140(11):3751-3757. (Immunogen: Flow cytometry, Fluorescence activated cell sorting, Immunofluorescence, Western blot). View Reference
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Hardy R, Hayakawa K. Generation of Ly-1 B cells from developmentally distinct precursors. Enrichment by stromal-cell culture or cell sorting. Ann N Y Acad Sci. 1992; 651:99-111. (Biology: Western blot). View Reference
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Hardy RR, Carmack CE, Shinton SA, Kemp JD, Hayakawa K. Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J Exp Med. 1991; 173(5):1213-1225. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Jones AT, Federsppiel B, Ellies LG, et al. Characterization of the activation-associated isoform of CD43 on murine T lymphocytes. J Immunol. 1994; 153(8):3426-3439. (Clone-specific: Western blot). View Reference
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Moore T, Bennett M, Kumar V. Transplantable NK cell progenitors in murine bone marrow. J Immunol. 1995; 154(4):1653-1663. (Clone-specific: Flow cytometry). View Reference
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Rolink A, ten Boekel E, Melchers F, Fearon DT, Krop I, Andersson J. A subpopulation of B220+ cells in murine bone marrow does not express CD19 and contains natural killer cell progenitors. J Exp Med. 1996; 183(1):187-194. (Clone-specific: Flow cytometry). View Reference
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Stockton BM, Cheng G, Manjunath N, Ardman B, von Andrian UH. Negative regulation of T cell homing by CD43. Immunity. 1998; 8(3):373-381. (Clone-specific: Flow cytometry). View Reference
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Thurman EC, Walker J, Jayaraman S, Manjunath N, Ardman B, Green JM. Regulation of in vitro and in vivo T cell activation by CD43. Int Immunol. 1998; 10(5):691-701. (Biology). View Reference
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Wells SM, Kantor AB, Stall AM. CD43 (S7) expression identifies peripheral B cell subsets. J Immunol. 1994; 153(12):5503-5515. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
771748 Rev. 1
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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.