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Flow cytometric analysis of CD43 expression on mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BUV737 Rat IgG2a, κ Isotype Control (Cat. No. 612760; dashed line histogram) or BD Horizon BUV737 Rat Anti-Mouse CD43 antibody (Cat. No. 612840; solid line histogram) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD43 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BUV737 Rat Anti-Mouse CD43
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. This will ensure that BD CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.
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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.