Overview

  • Product nameAnti-SSEA4 antibody [MC813]
  • Description
    Mouse monoclonal [MC813] to SSEA4
  • SpecificityThis antibody reacts with the Stage-specific embryonic antigen-4 (SSEA-4, a glycolipid carbohydrate epitope) that is expressed upon the surface of human teratocarcinoma stem cells (EC), human embryonic germ cells (EG) and human embryonic stem cells (ES). No immunoreactivity is evident with undifferentiated murine EC, ES and EG cells.Expression of SSEA-4 is down regulated following differentiation of human EC cells. In contrast, the differentiation of murine EC and ES cells may be accompanied by an increase in SSEA-4 expression.
  • Tested applicationsICC/IF, ICC, WB, Flow Cyt, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Human embryonal carcinoma cell line 2102Ep.

  • General notesThe mouse myeloma was fused with a spleen from a Balb/c mouse.

Properties

Applications

Our Abpromise guarantee covers the use of ab16287 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 15 µg/ml.
ICC Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
Flow Cyt Use a concentration of 15 µg/ml.
IHC-FoFr Use at an assay dependent concentration.

Target

  • RelevanceSSEA-4 (Stage-specific embryonic antigen 4) is a glycoprotein expressed early in embryonic development and in pluripotent stem cells. SSEA-4 can be used as a marker of Human Embryonic Stem Cells, Human Embryonic Carcinoma Cells and Human Embryonic Germ Cells. Monoclonal antibodies to this target have been widely used in the characterization of pluripotent stem cells. Mouse pluripotent stem cells are not recognised by anti-SSEA-4 antibodies but do express the antigen upon differentiation.
  • Cellular localizationCell Membrane
  • Alternative names
    • Stage specific embryonic antigen 4 antibody

Anti-SSEA4 antibody [MC813] images

  • ab16287 staining SSEA4 in rat tendon derived stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 in PBS and blocked with 1% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody (1/100) for 20 hours at 4°C. An Alexa Fluor® 594-conjugated donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

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  • ab16287 staining SSEA4 in human Fanconi's Anaemia iPS cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/300 in permeabilization buffer) for 1 hour at 4°C. An allophycocyanin-conjugated goat anti-mouse IgG3 polyclonal (1/800) was used as the secondary antibody.
    Gating Strategy: Isotype negative control (white).

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  • 2102ep Human Embryonic Carcinoma cells were stained with SSEA4 antibody ab16287. As expected, staining localised to the cell surface (green). Nuclei are stained blue using Hoechst.
  • anti-SSEA4 antibody, ab16297 can be used as a marker of Embryonic Carcinoma cells in Flow Cytometry/FACS. As can be seen from the histograms, in non-differentiating conditions (i.e. without retinoic acid) NTERA2 cells were recognised by ab16297. However, upon differentiation (addition of retinoic acid), the antibody lost the ability to recognise the cells.
  • ab16287 staining SSEA4 in human Embryonic Stem Cells, HUES7 by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with Triton and blocking with 10% serum for 1 hour was performed. Samples were incubated with primary antibody (1/100: in 1% serum, 0.1% Triton in PBS) for 1 hour at 37°C. An Alexa Fluor®588-conjugated goat polyclonal to mouse IgG was used at dilution at 1/100 as secondary antibody.

    See Abreview

  • Immunofluorescence analysis of Human dental pulp stem cells, staining SSEA4 with ab16287 at 1/40 dilution. A FITC-conjugated anti-mouse IgG was used as the secondary antibody.

References for Anti-SSEA4 antibody [MC813] (ab16287)

This product has been referenced in:
  • Sproul AA  et al. Generation of iPSC lines from archived non-cryoprotected biobanked dura mater. Acta Neuropathol Commun 2:4 (2014). ICC/IF ; Human . Read more (PubMed: 24398250) »
  • Begum AN  et al. Women with the Alzheimer's risk marker ApoE4 lose Aß-specific CD4? T cells 10-20 years before men. Transl Psychiatry 4:e414 (2014). Flow Cyt ; Human . Read more (PubMed: 25072319) »

See all 33 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Sample Rat Cell (tendon drived stem cell)
Specification tendon drived stem cell
Permeabilization Yes - PBS containing 0.25% Triton X-100
Fixative Paraformaldehyde
Username

Mrs. Sun Jeong Kim

Verified customer

Submitted Feb 25 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - eBioscience Permeabilization Buffer
Sample Human Cell (Fanconi's Anaemia iPSC)
Specification Fanconi's Anaemia iPSC
Gating Strategy Isotype negative control (white)
Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
Sample buffer: PBS and 10% FBS
Username

Abcam user community

Verified customer

Submitted Jan 28 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Sample Human Cell (iPS)
Specification iPS
Permeabilization No
Fixative Formaldehyde
Username

Mr. Faisal Alzahrani

Verified customer

Submitted Dec 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - 0.5% Saponin Buffer
Sample Human Cell (H9 Human Embryonic Stem Cells)
Specification H9 Human Embryonic Stem Cells
Gating Strategy FL2/FSC on isotype control
Preparation Cell harvesting/tissue preparation method: Single cell dissociation buffer
Sample buffer: FACS Buffer (5% FBS)
Username

Abcam user community

Verified customer

Submitted Jul 19 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (H9 embryonic stem cells)
Specification H9 embryonic stem cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Human Cultured Cells (H9 human)
Specification H9 human
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 10 2013

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization No
Sample Human Cell (human pluripotent iPS cells)
Specification human pluripotent iPS cells
Gating Strategy FSC vs SSC on negative control sample
Preparation Cell harvesting/tissue preparation method: cell dissociation buffer
Sample buffer: FACS buffer (0.2% BSA, 0.1% sodium azide in PBS)
Username

Abcam user community

Verified customer

Submitted Jul 04 2013

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Sample Human Cultured Cells (induced pluripotent stem cells)
Specification induced pluripotent stem cells
Permeabilization No
Fixative Methanol
Username

Abcam user community

Verified customer

Submitted Jun 07 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Sample Human Cell (Embryonic Stem Cell)
Specification Embryonic Stem Cell
Permeabilization Yes - BD Perm/Wash Buffer
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 21 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (Embryonic Stem Cell)
Specification Embryonic Stem Cell
Preparation Cell harvesting/tissue preparation method: Trypsinisation
Sample buffer: BD Perm/Wash Buffer
Fixation Paraformaldehyde
Permeabilization Yes - BD Perm/Wash Buffer
Gating Strategy Gating position dependent on isotype control population
Username

Abcam user community

Verified customer

Submitted May 15 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"