Two-color flow cytometric analysis of Ly-76 (TER-119) expression on Mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) antibody (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45 antibody (Cat. No. 559864) and with either no antibody (Autofluorescence Control; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse Ly-76 (TER-119) antibody (Cat. No. 569365/569366; Right Plot) 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 bivariate pseudocolor density plot showing the correlated expression of Ly-76 (TER-119) [or Autofluorescence] versus CD45 were 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 LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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Two-color flow cytometric analysis of Ly-76 (TER-119) expression on Mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) antibody (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45 antibody (Cat. No. 559864) and with either no antibody (Autofluorescence Control; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse Ly-76 (TER-119) antibody (Cat. No. 569365/569366; Right Plot) 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 bivariate pseudocolor density plot showing the correlated expression of Ly-76 (TER-119) [or Autofluorescence] versus CD45 were 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 LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
Lymphocyte antigen 76; Ly76; Ly-76; TER-119; Ter119
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2b, κ
Mouse Fetal Liver
Flow cytometry (Routinely Tested)
0.2 mg/ml
104231
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 and unreacted dye was removed.
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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
APC Rat Anti-Mouse CD45 RUO
Size
0.1 mg
Cat No.
559864
569365 Rev. 2
Antibody Details
TER-119
The TER-119 antibody specifically binds to a 52 kDa molecule associated with glycophorin A on cells of the erythroid lineage in embryonic yolk sac, fetal liver, newborn liver, adult bone marrow, adult peripheral blood, and adult lymphoid organs. The TER-119 antigen is expressed on erythroid cells from pro-erythroblast through mature erythrocyte stages, but not on cells with BFU-E or CFU-E activities. The TER-119 epitope is not detected on hematopoietic stem cells, lymphoid cells, myeloid cells, or erythroleukemia lines. The TER-119 mAb is a component of the "lineage cocktail" used in studies of hematopoietic progenitors to detect, or deplete cells committed to the hematopoietic lineages.
569365 Rev. 2
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
569365 Rev.2
Citations & References
View product citations for antibody "569365" on CiteAb
Development References (5)
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Ikuta K, Kina T, MacNeil I, et al. A developmental switch in thymic lymphocyte maturation potential occurs at the level of hematopoietic stem cells. Cell. 1990; 62(5):863-874. (Clone-specific). View Reference
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Kina T, Ikuta K, Takayama E, et al. The monoclonal antibody TER-119 recognizes a molecule associated with glycophorin A and specifically marks the late stages of murine erythroid lineage. Br J Haematol. 2000; 109(2):280-287. (Immunogen: Immunoprecipitation, Western blot). View Reference
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Kitajima K, Kojima M, Nakajima K, et al. Definitive but not primitive hematopoiesis is impaired in jumonji mutant mice. Blood. 1999; 93(1):87-95. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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Maraskovsky E, Brasel K, Teepe M, et al. Dramatic increase in the numbers of functionally mature dendritic cells in Flt3 ligand-treated mice: multiple dendritic cell subpopulations identified. J Exp Med. 1996; 184(5):1953-1962. (Clone-specific: Cell separation, Cytotoxicity, Depletion, Flow cytometry). View Reference
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Osawa M, Tokumoto Y, Nakauchi H. Hematopoietic stem cells. In: Herzenberg LA, Weir DM, Blackwell C, ed. Weir's Handbook of Experimental Immunology, 5th Edition. Cambridge: Blackwell Science; 1996:66.1-66.5.
569365 Rev. 2
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.