Flow cytometric analysis of CD43 expression on mouse bone marrow cells. BALB/c 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 PE-Cy™7 Rat Anti-Mouse CD43 antibody (Cat. No. 562866, solid line histogram) or with PE-Cy™7 Rat IgG2a, κ Isotype Control (Cat. No. 552784, dashed line histogram). Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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Flow cytometric analysis of CD43 expression on mouse bone marrow cells. BALB/c 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 PE-Cy™7 Rat Anti-Mouse CD43 antibody (Cat. No. 562866, solid line histogram) or with PE-Cy™7 Rat IgG2a, κ Isotype Control (Cat. No. 552784, dashed line histogram). Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Spn; Sialophorin; Leukosialin; Ly-48; Ly48; Galgp; LEUK
Mouse (QC Testing)
Rat DA x LOU IgG2a, κ
Mouse Plasmacytoma MOPC-315
Flow cytometry (Routinely Tested)
0.2 mg/ml
20737
AB_2737852
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 with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- 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.
- 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.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. 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 PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
PE-Cy™7 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
552784
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562866 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.
562866 Rev. 1
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
562866 Rev.1
Citations & References
Development References (9)
-
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
562866 Rev. 1
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.
