SSEA-3 Antibody (MC-631) - BSA Free

Novus Biologicals | Catalog # NB100-1832

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, CyTOF-ready

Cited:

Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Rat IgM Clone # MC-631

Format

BSA Free
View all SSEA-3 Primary Antibodies »

Product Specifications

Immunogen

4-8 cell stage mouse embryos.

Reactivity Notes

This antibody recognizes the SSEA-3 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-3 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-3 expression.

Marker

Embryonic Stem Cell Marker

Specificity

Recognizes a carbohydrate epitope of SSEA-3

Clonality

Monoclonal

Host

Rat

Isotype

IgM

Scientific Data Images for SSEA-3 Antibody (MC-631) - BSA Free

Immunocytochemistry/ Immunofluorescence: SSEA-3 Antibody (MC-631) [NB100-1832]

Immunocytochemistry/ Immunofluorescence: SSEA-3 Antibody (MC-631) [NB100-1832]

SSEA-3-Antibody-MC-631-Immunocytochemistry-Immunofluorescence-NB100-1832-img0016.jpg
Flow Cytometry: SSEA-3 Antibody (MC-631) [NB100-1832]

Flow Cytometry: SSEA-3 Antibody (MC-631) [NB100-1832]

Flow Cytometry: SSEA-3 Antibody (MC-631) [NB100-1832] - A surface stain was performed on Jurkat cells with SSEA-3 Antibody (MC-631) NB100-1832AF647 (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Immunocytochemistry/ Immunofluorescence: SSEA-3 Antibody (MC-631) [NB100-1832]

Immunocytochemistry/ Immunofluorescence: SSEA-3 Antibody (MC-631) [NB100-1832]

Immunocytochemistry/Immunofluorescence: SSEA-3 Antibody (MC-631) [NB100-1832] - Analysis of human embryonic stem cells stained with SSEA-3 antibody detected with Alexa Fluor 488 anti-rat IgM secondary antibody.
SSEA-3 Antibody (MC-631) - BSA Free

Immunocytochemistry/ Immunofluorescence: SSEA-3 Antibody (MC-631) - BSA Free [NB100-1832] -

PSC characterisation. (A) Human PSC-1 characterized for A) typical undifferentiated colony morphology in phase contrast image and (B) expression of pluripotency markers Nanog, OCT-3/4, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81, and lack of expression of early differentiation marker SSEA-1 after immunofluorescence staining. Corresponding nuclei stains with DAPI shown. (C) Cells showed normal female (46, XX) karyotype after 28 passages in total (9 passages in feeder-free culture). The results of the KaryoLite BoBs assay are shown as signal relative to karyotypically normal female (/F, red) and male (/M, blue) genomic DNA used as a reference (equal to 1) for each of the probes covering both p and q arms of all chromosomes. Software threshold for changes shown as a green lines and deviations in red. (D) Pluripotency shown after spontaneous differentiation in vitro as expression of markers for mesoderm, endoderm, and ectoderm. All scale bars 200 um. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30341351), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for SSEA-3 Antibody (MC-631) - BSA Free

Application
Recommended Usage

Flow Cytometry

10-20 ug/ml

Immunocytochemistry/ Immunofluorescence

1:50-1:100

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:10-1:500

Western Blot

1:500
Application Notes
This SSEA3 antibody is useful for Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry frozen and paraffin sections, Immunoprecipitation and Western Blot. This antibody is CyTOF ready.

Reviewed Applications

Read 1 review rated 5 using NB100-1832 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

IgM purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: SSEA-3

SSEA-3 (stage-specific embryonic antigen-3) is a member of SSEAs which are present on eggs as well as preimplantation embryos at specific stages of early development. SSEAs are glycan epitopes which resides on glycoproteins/glycolipids of mammalian cells, and the epitope of several of these antigens consists solely of sugar residues. SSEA-3 antigen was identified following immunization of rats with 4-8 cell mouse embryos and was present on glycolipids of human teratocarcinoma cells. During protein glycosylation, beta-1,3-galactosyltransferase 5 catalyzes the transfer of Gal to the terminal GalNAc unit of the globoside GB4, thereby synthesizing glycolipid GB5 which is SSEA-3. In its functions. SSEA-3 has been proposed to involve in the process of cell differentiation. SSEA3 expression is mostly restricted to the cytoplasm or apical surface of epithelial cells and it has been reported to express in various normal tissues with high expression levels in kidney. SSEA3 has been reported to be expressed in the ascites fluids from patients of hepatoma or pancreatic cancer, breast cancer including breast cancer stem cells.

Long Name

Stage-specific Embryonic Antigen-3

Alternate Names

SSEA3

Additional SSEA-3 Products

Product Documents for SSEA-3 Antibody (MC-631) - BSA Free

Certificate of Analysis

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Product Specific Notices for SSEA-3 Antibody (MC-631) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for SSEA-3 Antibody (MC-631) - BSA Free

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  • Name: Gail M. Seigel
    Application: Immunohistochemistry-Paraffin
    Sample Tested: HeLa cells
    Species: Human
    Verified Customer | Posted 08/07/2010
    SSEA-3 Antibody (MC-631) - BSA Free NB100-1832

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Protocols

View specific protocols for SSEA-3 Antibody (MC-631) - BSA Free (NB100-1832):

SSEA-3 Antibody (MC-631):
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for SSEA-3 Antibody (MC-631) - BSA Free

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  • Q: Could you please let us know the accession number of the antigen protein for SSEA3 Antibody (Cat. No. NB100-1832)?

    A: SSEA3 is a globoseries carbohydrate antigen found on glycolipids as well as glycopeptides on cellular surface of embryonic stem cells. Because it is not a protein in reality, it is not supposed to have an accession number. NB100-1832 was generated in rat host using murine 4-8 cell stage embryos as immunogen and this antibody specifically detects the SSEA3 antigen in various immunoassays.

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