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PerCP-Cy™5.5 Mouse Anti-SSEA-1
PerCP-Cy™5.5  Mouse Anti-SSEA-1
Flow cytometric analysis of SSEA-1 expression on E14 cells.   E14 mouse embryonic stem (ES) cells were stained with either PerCP-Cy™5.5 Mouse Anti-SSEA-1 (Cat. No. 561560; solid line histogram) or PerCP-Cy™5.5 Mouse IgM, κ Isotype Control (Cat. No. 560857, dashed line histogram), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Fluorescence histogram depicting SSEA-1 (or Ig isotype expression) were derived from gated events with the side and forward light-scattering characteristics of viable cells. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of SSEA-1 expression on E14 cells.   E14 mouse embryonic stem (ES) cells were stained with either PerCP-Cy™5.5 Mouse Anti-SSEA-1 (Cat. No. 561560; solid line histogram) or PerCP-Cy™5.5 Mouse IgM, κ Isotype Control (Cat. No. 560857, dashed line histogram), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Fluorescence histogram depicting SSEA-1 (or Ig isotype expression) were derived from gated events with the side and forward light-scattering characteristics of viable cells. Flow cytometry was performed on a BD FACSCalibur™ System.
製品詳細
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BD Pharmingen™
3-FAL, X-hapten, LeX antigen, CD15
Mouse (QC Testing), Human (Tested in Development)
Mouse BALB/c IgM, κ
Mouse Teratocarcinoma Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_11153302
Aqueous buffered solution containing BSA, protein stabilizer, and ≤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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

推奨アッセイ手順

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561560 Rev. 3
抗体の詳細
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MC480

The MC480 monoclonal antibody reacts with Stage-Specific Embryonic Antigen-1 (SSEA-1), which is a terminal carbohydrate epitope (3-fucosyl-N-acetyllactosamine or 3-FAL) on glycoproteins and lactose-series glycolipids.  SSEA-1 is related to Lewis blood group antigens and is found in a variety of embryonic as well as adult tissues and cancers.  As its name implies, the expression of SSEA-1 is stage-specific and can be used to characterize embryonic cells and monitor their differentiation.  However, its expression pattern differs between human and mice.  In the human, SSEA-1 is not found on embryonic stem (ES) cells, embryonic inner cell mass (ICM), or teratocarcinoma (embryonal carcinoma or EC) cells.  As human EC and ES cells undergo differentiation, SSEA-1 expression is upregulated.  In the adult, the same epitope is expressed as CD15 on granulocytes and monocytes, but not lymphocytes or dendritic cells.  In the mouse, SSEA-1 is found on EC, ES, primordial germ cells, 8-cell to blastocyst embryos, ICM, and subpopulations of cells in the adult central nervous system, including stem cells.  In contrast to human SSEA-1 expression, it is reduced when mouse EC and ES cells undergo differentiation.

561560 Rev. 3
フォーマットの詳細
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
561560 Rev.3
引用&参考文献
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Development References (7)

  1. Capela A, Temple S. LeX/ssea-1 is expressed by adult mouse CNS stem cells, identifying them as nonependymal. Neuron. 2002; 35:865-875. (Biology).
  2. Childs RA, Pennington J, Uemura K, et al. High-molecular-weight glycoproteins are the major carriers of the carbohydrate differentiation antigens I, i and SSEA-1 of mouse teratocarcinoma cells. Biochem J. 1983; 215:491-503. (Clone-specific).
  3. Draper JS, Pigott C, Thomson JA, Andrews PW. Surface antigens of human embryonic stem cells: changes upon differentiation in culture. J Anat. 2002; 200:249-258. (Clone-specific). View Reference
  4. Henderson JK, Draper JS, Baillie HS, et al. Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens. Stem Cells. 2002; 20:329-337. (Clone-specific). View Reference
  5. Kannagi R, Nudelman E, Levery SB, Hakomori S. A series of human erythrocyte glycosphingolipids reacting to the monoclonal antibody directed to a developmentally regulated antigen, SSEA-1. J Biol Chem. 1982; 257(24):14865-14874. (Clone-specific).
  6. Solter D, Knowles BB. Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1). Proc Natl Acad Sci U S A. 1978; 75(11):5565-5569. (Immunogen). View Reference
  7. Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282:1145-1147. (Clone-specific). View Reference
すべて表示する (7) 表示項目を減らす
561560 Rev. 3

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