Clone:
REA101
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC

Extended validation for SSEA-4 Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA101
MC813++
Cells were incubated with an excess of purified unconjugated SSEA-4 (REA101) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for SSEA-4. NTERA-2 cells were stained with SSEA-4 antibodies and plotted against the side scatter. As a control, SSEA-4 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for SSEA-4. NTERA-2 cells were stained with SSEA-4 antibodies and plotted against the side scatter. As a control, SSEA-4 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for SSEA-4. NTERA-2 cells were stained with SSEA-4 antibodies and plotted against the side scatter. As a control, SSEA-4 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for SSEA-4. NTERA-2 cells were stained with SSEA-4 antibodies and plotted against the side scatter. As a control, SSEA-4 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using SSEA-4 (REA101). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using SSEA-4 (REA101). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using SSEA-4 (REA101). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for SSEA-4 Antibody, anti-human, REAfinity™

Overview

Clone REA101 recognizes the glycolipid carbohydrate epitope stage specific embryonic antigen 4 (SSEA-4). SSEA-4 is found on undifferentiated human embryonic stem (ES) cells, induced pluripotent (iPS) cells, embryonal carcinoma (EC) cells, as well as embryonic germ (EG) cells and a variety of somatic stem cells, such as dental pulp stem cells, umbilical cord blood–derived very small embryonic like stem cells (VSELs), and mesenchymal stromal cells.
Additional information: Clone REA101 displays negligible binding to Fc receptors.

Detailed product information

Technical specifications

CloneREA101
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse, other
Cross-reactivitymouse, rabbit, canine, chicken
AntigenSSEA-4
Distribution of antigenstem cells, ES and iPS cells
RRIDAB_2811405, AB_2811416, AB_2811404, AB_2877030, AB_2877031, AB_2876944, AB_2876945, AB_2653521, AB_2653528, AB_2811417

Resources for SSEA-4 Antibody, anti-human, REAfinity™

Certificates

Please follow this
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to search for Certificates of Analysis (CoA) by lot number.

References for SSEA-4 Antibody, anti-human, REAfinity™

Publications

  1. Gang, E. J. et al. (2007) SSEA-4 identifies mesenchymal stem cells from bone marrow. Blood 109(4): 1743-1751
  2. Kannagi, R. et al. (1983) Stage-specific embryonic antigens (SSEA-3 and -4) are epitopes of a unique globo-series ganglioside EMBO J. 2(12): 2355-2361
  3. Truong, T. T. et al. (2011) SSEA4 is a potential negative marker for the enrichment of human corneal epithelial stem/progenitor cells. Invest. Ophthalmol. Vis. Sci. 52(9): 6315-6320
  4. Aloia, A. et al. (2015) The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features. Breast Cancer Res. 17: 146

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