Testing Data
(Image: SSEA3 staining of HeLa cells (Green) detected with Fluor 488 conjugated anti-rat IgM secondary antibody. Nuclei are counterstained with Hoechst 33258 (Blue). Protocol on Datasheet.)
Rat anti-Human, Mouse SSEA3 Antibody | anti-SSEA3 antibody
SSEA3, Affinity Purified Antibody, rat
Reactivity
Human, Mouse
Applications
Immunocytochemistry, Western Blot, Flow Cytometry, Functional Assay, Immunoprecipitation
Synonyms
SSEA3; Affinity Purified Antibody; rat; anti-SSEA3 antibody
Host
Rat
Reactivity
Human, Mouse
Specificity
This antibody reacts with the Stage-specific embryonic antigen-3 (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
Form/Format
Liquid. Tris-glycine, 150mM NaCl with 0.05% Sodium Azide as a preservative.
Concentration
1 mg/ml. (varies by lot)
Applicable Applications for anti-SSEA3 antibody
Immunocytochemistry (ICC), Western Blot (WB), Flow Cytometry (FC/FACS), Immunoprecipitation (IP)
Application Notes
Immunocytochemistry 1:50 -1:100
Flow Cytometry: 1:50
Immunocytochemistry:
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.
Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.
Flow Cytometry: 1:50
Immunocytochemistry:
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.
Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.
Immunogen
4-8 cell stage mouse embryos.
Preparation and Storage
Store frozen. Aliquot as undiluted antisera and immediately place at -20°C. Antisera may have become trapped in top of vial during shipping. Centrifugation of vial is recommended before opening. Stable for at least 6 months at -20°C. Repeated freeze/thaw cycles compromise the integrity of the antiserum.
Testing Data
(Image: SSEA3 staining of HeLa cells (Green) detected with Fluor 488 conjugated anti-rat IgM secondary antibody. Nuclei are counterstained with Hoechst 33258 (Blue). Protocol on Datasheet.)
Testing Data
(Image: SSEA3 staining of HeLa cells (Green) detected with Fluor 488 conjugated anti-rat IgM secondary antibody. Nuclei are counterstained with Hoechst 33258 (Blue). Protocol on Datasheet.)
Testing Data
(Image: FACS staining of NTERA-2 cells using SSEA3 at a 1:50 dilution detected using 488 conjugated goat anti-rat IgM secondary antibody.)
Testing Data
(Image: FACS staining of NTERA-2 cells using SSEA3 at a 1:50 dilution detected using 488 conjugated goat anti-rat IgM secondary antibody.)
Related Product Information for anti-SSEA3 antibody
SSEA3 (stage-specific mouse embryonic antigen) is a cell-surface protein involved in cell differentiation. Cell surface antigens are powerful tools for characterizing the function and expression of developmentally regulated cells during both embryogenesis and oncogenesis.
Product Categories/Family for anti-SSEA3 antibody
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Product Notes
The SSEA3 (Catalog #AAA555001) is an Antibody produced from Rat and is intended for research purposes only. The product is available for immediate purchase. The SSEA3, Affinity Purified Antibody, rat reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's SSEA3 can be used in a range of immunoassay formats including, but not limited to, Immunocytochemistry (ICC), Western Blot (WB), Flow Cytometry (FC/FACS), Immunoprecipitation (IP). Immunocytochemistry 1:50 -1:100 Flow Cytometry: 1:50 Immunocytochemistry: 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. Dilutions listed as a recommendation. Optimal dilution should be determined by investigator. Researchers should empirically determine the suitability of the SSEA3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "SSEA3, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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