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RB705 Rat Anti-Mouse Ly-6A/E (Sca-1)
Product Details
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BD OptiBuild™
SCA-1; Sca1; Stem cell antigen 1; TAP; T-cell-activating protein; Ly6
Mouse (Tested in Development)
Rat WI, also known as Wistar (outbred) IgG2a, κ
BALB/c mouse-derived "pre-T" cell hybridoma
Flow cytometry (Qualified)
0.2 mg/ml
110454
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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. 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.
757750 Rev. 1
Antibody Details
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E13-161.7

The E13-161.7 monoclonal antibody recognizes Ly-6A.2 and Ly-6E.1, which are allelic members of the Ly-6 multigene family. Ly-6A/E is also known as, stem cell antigen 1 (Sca-1/Sca1), or T-cell-activating protein  (TAP). Ly-6A/E is a phosphatidylinositol-anchored protein of ~18 kDa that is expressed on multipotent hematopoietic stem cells (HSC) in mice with both Ly-6 haplotypes. Sca-1+ HSC are found in the adult bone marrow and fetal liver, but not in the early embryo yolk sac or intraembryonic hematopoietic sites, and can be mobilized to the peripheral blood and spleen in the adult. In mice expressing the Ly-6.2 haplotype (e.g., AKR, C57BL, C57BR, C57L, C58, DBA/2, PL, SJL, SWR, 129), Ly-6A/E is also expressed on distinct subpopulations of bone marrow and peripheral B lymphocytes, myeloid cells, and thymic and peripheral T lymphocytes, on the earliest intrathymic T-cell precursor population, and in several non-hematopoietic tissues. Strains with the Ly-6.1 haplotype (e.g., A, BALB/c, CBA, C3H/He, DBA/1, NZB) have few Ly-6A/E+ resting peripheral lymphocytes, whereas activation of T cells from mice of both Ly-6 haplotypes leads to strong expression of the Sca-1 antigen. Studies with the D7 antibody have demonstrated that Ly-6A/E may be involved in the regulation of B and T lymphocyte responses, and it appears to be required for T-cell receptor-mediated T-cell activation. Purified E13-161.7 mAb can block binding of FITC-conjugated D7 antibody (anti-Ly-6A/E) to mouse splenocytes, but purified mAb D7 is unable to block binding of FITC-conjugated E13-161.7 antibody. Anti-Ly-6A/E (Sca-1) mAb may be used in combination with a Mouse Lineage Antibody Panel (e.g., Cat. No. 559971) to identify HSC.

757750 Rev. 1
Format Details
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RB705
The BD Horizon RealBlue™ 705 (RB705) 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 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 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), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
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RB705
Blue 488 nm
498 nm
707 nm
757750 Rev.1
Citations & References
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View product citations for antibody "757750" on CiteAb

Development References (4)

  1. Aihara Y, Buhring HJ, Aihara M, Klein J. An attempt to produce "pre-T" cell hybridomas and to identify their antigens. Eur J Immunol. 1986; 16(11):1391-1399. (Immunogen: Cytotoxicity, Flow cytometry). View Reference
  2. Kawamoto H, Ohmura K, Katsura Y. Direct evidence for the commitment of hematopoietic stem cells to T, B and myeloid lineages in murine fetal liver. Int Immunol. 1997; 9(7):1011-1019. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  3. Marcos MA, Morales-Alcelay S, Godin IE, Dieterlen-Lievre F, Copin SG, Gaspar ML. Antigenic phenotype and gene expression pattern of lymphohemopoietic progenitors during early mouse ontogeny. J Immunol. 1997; 158(6):2627-2637. (Biology). View Reference
  4. Osawa M, Nakamura K, Nishi N, et al. In vivo self-renewal of c-Kit+ Sca-1+ Lin(low/-) hemopoietic stem cells. J Immunol. 1996; 156(9):3207-3214. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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757750 Rev. 1

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.