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APC Mouse Anti-SSEA-1
APC Mouse Anti-SSEA-1
Flow cytometric analysis of SSEA-1 expression on mouse embryonic stem (ES) cells.  ES-E14TG2a mouse ES cells (ATCC, CRL-1821™) at passage 40 were harvested with Accutase™ Cell detachment solution (Cat. No. 561527) and preserved with BD Cytofix™ fixation buffer (Cat. No. 554655). The cells were washed with stain buffer before staining with either APC Mouse Anti-SSEA-1 (Cat. No. 566790, solid line histogram) or APC Mouse IgM, κ Isotype Control (Cat. No.550883, dashed line histogram). The fluorescence histogram showing SSEA-1 (or Ig Isotype control) staining was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD FACS LSRFortessa™ X20 flow cytometer and FlowJo™ software.
Flow cytometric analysis of SSEA-1 expression on mouse embryonic stem (ES) cells.  ES-E14TG2a mouse ES cells (ATCC, CRL-1821™) at passage 40 were harvested with Accutase™ Cell detachment solution (Cat. No. 561527) and preserved with BD Cytofix™ fixation buffer (Cat. No. 554655). The cells were washed with stain buffer before staining with either APC Mouse Anti-SSEA-1 (Cat. No. 566790, solid line histogram) or APC Mouse IgM, κ Isotype Control (Cat. No.550883, dashed line histogram). The fluorescence histogram showing SSEA-1 (or Ig Isotype control) staining was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD FACS LSRFortessa™ X20 flow cytometer and FlowJo™ software.
Product Details
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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_2869868
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 to APC under optimum conditions, and unconjugated antibody and free APC were removed.

Recommended Assay Procedures

Note: For best staining results, cell samples should be stained and maintained at 4-8°C before flow cytometric analysis.

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 cells 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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566790 Rev. 2
Antibody Details
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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.

566790 Rev. 2
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
566790 Rev.2
Citations & References
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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: Immunofluorescence, Western blot).
  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: Flow cytometry). 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: Flow cytometry, Immunofluorescence). 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: Cytotoxicity, Radioimmunoassay). 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: Immunocytochemistry (cytospins)). View Reference
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566790 Rev. 2

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