Stage Specific Embryonic Antigen 3 (SSEA3) antibody

Details for Product No. ABIN151342
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Antigen
Reactivity
Human, Mouse (Murine)
(15), (14), (12)
Host
Rat
(12), (4)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(10), (4), (4), (3), (2), (2), (1), (1), (1)
Pubmed 1 reference available
Quantity 0.1 mL
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Catalog No. ABIN151342
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Immunogen 4-8 cell stage mouse embryos.
Clone MC-631
Isotype IgM
Specificity This antibody recognizes the SSEA3 that is expressed upon the surface of human teratocarcinoma stem cells (EC), human embryonic germ cells (EG),human embryonic stem cells (ES). No immunoreactivity is evident with undifferentiated murine EC, ES,EG cells. Expression of SSEA3 is down regulated following differentiation of human EC cells. In contrast, the differentiation of murine EC,ES cells may be accompanied by an increase in SSEA-3 expression.
Purification IgM purified
Alternative Name SSEA3
Background SSEA3 (stage-specific mouse embryonic antigen) is a cell-surface protein involved incell differentiation. Cell surface antigens are powerful tools for characterizing thefunction and expression of developmentally regulated cells during both embryogenesisand oncogenesis. Alternate Names: Anti-Stage specific embryonic antigen 3 antibody, Anti-Stage-specific embryonic antigen3 antibody.
Application Notes This SSEA3 antibody is useful for Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry frozen and paraffin sections, Immunoprecipitation and Western Blot.
Recommended dilutions: Flow Cytometry 10-20 µg/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
Protocol Protocol Specific for NB100-1832 Immunocytochemistry ProtocolCulture 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.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. 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.
. 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.
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer Tris-glycine, 150 mM NaCl, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
General Henderson, Draper, Baillie et al.: "Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens." in: Stem cells (Dayton, Ohio), Vol. 20, Issue 4, pp. 329-37, 2002 (PubMed).

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