Overview

  • Product name
    Anti-SSEA4 antibody [MC813]
  • Description
    Mouse monoclonal [MC813] to SSEA4
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cyt, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Human embryonal carcinoma cell line 2102Ep.

  • General notes

    This antibody clone is manufactured by Abcam.

    The 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.
WB Use at an assay dependent concentration.
Flow Cyt Use a concentration of 15 µg/ml.

ab18392 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.

IHC-FoFr Use at an assay dependent concentration.

Target

  • Relevance
    Glycosphingolipids function as mediators of cell adhesion and are modulators of signal transduction. SSEA-4 (Stage-specific embryonic antigen 4) is a glycolipid 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 localization
    Cell 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.

    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.
  • Immunocytochemistry/ Immunofluorescence analysis of human iPSC cells labeling SSEA4 with ab16287 at 1/500 dilution. Cells were fixed with paraformaldehyde and  permeabilized with 0.5% TX100. Cells were blocked with 5% serum for 20 minutes at 25°C. A goat polyclonal anti-mouse Cy3 secondary antibody at 1/500 dilution was used. 

    See Abreview

  • 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.
  • 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.
  • 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

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

This product has been referenced in:
  • Li Y  et al. Abnormal Neural Progenitor Cells Differentiated from Induced Pluripotent Stem Cells Partially Mimicked Development of TSC2 Neurological Abnormalities. Stem Cell Reports 8:883-893 (2017). IF ; Human . Read more (PubMed: 28344003) »
  • Niu X  et al. Combining Single Strand Oligodeoxynucleotides and CRISPR/Cas9 to Correct Gene Mutations in ß-Thalassemia-induced Pluripotent Stem Cells. J Biol Chem 291:16576-85 (2016). ICC/IF ; Human . Read more (PubMed: 27288406) »

See all 45 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human iPSC cells)
Permeabilization
Yes - 0.5% TX100
Specification
human iPSC cells
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 23 2016

Application
Flow Cytometry
Sample
Human Cell (HEK 293)
Permeabilization
No
Gating Strategy
FSC vs SSC on negative control sample
Specification
HEK 293
Preparation
Cell harvesting/tissue preparation method: Trypsin
Sample buffer: 0.1% BSA in PBS -Ca-Mg
Fixation
no fixation
Username

Dr. Tove Berg

Verified customer

Submitted Dec 21 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (testis tissues)
Specification
testis tissues
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Mr. Heejun Jung

Verified customer

Submitted Aug 19 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (iCell® iPSC Diff Day 0)
Permeabilization
No
Specification
iCell® iPSC Diff Day 0
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Aug 11 2015

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
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
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

Thank you for contacting us.
I can confirm that the anti-CD24 antibody (ab110448), anti-CD44 antibody (ab119863), and anti-SSEA4 antibody (ab16287) are not conjugated to any fluorophores. A secondary antibody needs to be used.
I found in our p...

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