Ssea 1

Manufactured by Thermo Fisher Scientific

Sourced in United States

SSEA-1 is a cell surface antigen that is commonly used as a marker for embryonic stem cells. It is a carbohydrate epitope expressed on the surface of undifferentiated embryonic stem cells and some other cell types. The SSEA-1 antibody can be used to identify and isolate SSEA-1 positive cells.

Automatically generated - may contain errors

Lab products found in correlation

8 protocols using ssea 1

Protein Expression Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

On the day of the experiment, equal amounts of protein sample were loaded on an 8-10% sodium dodecyl sulfate (SDS) gel and separated using SDS polyacrylamide gel electrophoresis (SDS-PAGE). Then, the protein samples were transferred to a polyvinylidene difluoride (PVDF) membrane for 1–2 hours at 100 V. After a blocking step with PBS containing 3% BSA, the transferred membrane was incubated with primary antibody. The following day, the membrane was incubated with horseradish peroxidase (HRP)-conjugated secondary antibody for 1 hour, and then the detection procedure was performed. Primary antibodies were: stage-specific embryonic antigen 1 (SSEA-1; 1:1000, Invitrogen), actin (1:10000, Abcam), and C-Myc (1:500, Santacruz).

Oh J., Kim Y., Che L., Kim J.B., Chang G.E., Cheong E., Kang S.G, & Ha Y. (2017). Regulation of cAMP and GSK3 signaling pathways contributes to the neuronal conversion of glioma. PLoS ONE, 12(11), e0178881.

+ Open protocol

+ Expand

Immunostaining Induced Pluripotent Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

iPSCs were fixed in 4% paraformaldehyde solution for 15 min at room temperature (RT). Then they were permeabilized by using 0.1% Triton X-100 in PBS for 20 min at RT. After they were blocked in 5% goat serum for 1 h, the cells were incubated overnight at 4 °C by primary antibodies, including Nanog (Abcam), SSEA-1 (Invitrogen), Oct4 (Cell signal), SSEA-4 (ThermoFisher), Sox2 (Epitomics) and TRA-1-60 (Abcam). Finally, these cells were incubated by fluorescently coupled secondary antibodies (Invitrogen) and 4′,6-diamidino-2-phenylindole (DAPI; Sigma) for 1 h at RT. Images were captured on a confocal microscope (Leica).

Zou H., Guan M., Li Y., Luo F., Wang W, & Qin Y. (2021). Targeted gene correction and functional recovery in achondroplasia patient-derived iPSCs. Stem Cell Research & Therapy, 12, 485.

+ Open protocol

+ Expand

iPSC Characterization by Immunofluorescence

Check if the same lab product or an alternative is used in the 5 most similar protocols

The iPSC clones were confirmed as IPSCs with a battery of rabbit anti-mouse IPSC markers including Oct3/4, Sox2, c-Myc, mKlf4, Nestin and SSEA-1 (Thermo Fisher Scientific, Waltham, MA, USA). The secondary antibody was an Alexa Fluor 594-conjugated goat anti-rabbit (Thermo Fisher Scientific), all used in accordance with the manufacturer’s conditions. To immobilize the iPSC clones, glass-bottom dishes were coated with Cell-TEK adhesive. The adherent iPSC was then fixed with 4% paraformaldehyde, after permeabilizing with TX-100 and blocking with normal goat serum. The iPSC clones were then incubated with the primary antibodies, washed, and incubated with the secondary antibodies. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories, Burlingame, CA, USA) and viewed with an Olympus Fluoview-1000 confocal scanning system under different wavelengths.

Petrova S.C., Ahmad I., Nguyen C., Ferrell SD J.r., Wilhelm S.R., Ye Y, & Barsky S.H. (2020). Regulation of breast cancer oncogenesis by the cell of origin’s differentiation state. Oncotarget, 11(43), 3832-3848.

+ Open protocol

+ Expand

Immunofluorescence Staining of Stem Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were fixed in 4% paraformaldehyde (Sigma-Aldrich). After permeabilization with 0.1% Triton X-100 (Sigma-Aldrich), they were incubated with appropriate blocking solution (10% normal goat serum) followed by primary antibodies against SSEA-1 (Thermo Fisher Scientific), OCT4 (Santa Cruz Biotechnology), NANOG (Millipore), γH2AX (BD Biosciences, California, USA), WRN (Abcam) and 53BP1 (Santa Cruz Biotechnology) at 4°C overnight. The cells were then incubated with fluorescent-conjugated secondary antibodies (Thermo Fisher Scientific). The nucleus was stained with Hoechst 33258 (Thermo Fisher Scientific). Images of the staining cells were captured using a confocal microscope (LSM 700, Carl Zeiss AG, Oberkochen, Germany) at the Faculty Core Facility, The University of Hong Kong.

Chen A.C., Peng Q., Fong S.W., Yeung W.S, & Lee Y.L. (2020). Sirt1 is regulated by miR-135a and involved in DNA damage repair during mouse cellular reprogramming. Aging (Albany NY), 12(8), 7431-7447.

+ Open protocol

+ Expand

Characterization of Mouse iPSC Clones

Check if the same lab product or an alternative is used in the 5 most similar protocols

The iPSC clones were confirmed as IPSCs with a battery of rabbit anti-mouse IPSC markers including Oct3/4, Sox2, c-Myc, mKlf4, nestin, and SSEA-1 (Thermo Fisher Scientific, Waltham, MA) The secondary antibody was an Alexa Fluor 594-conjugated goat anti-rabbit (Thermo Fisher Scientific), all used with the manufacturer’s conditions. In order to immobilize the iPSC clones, glass-bottom dishes were coated with Cell-TEK adhesive. The adherent iPSC cells were then fixed with 4% paraformaldehyde, after permeabilizing with TX-100 and blocking with normal goat serum. The iPSC clones were then incubated with the primary antibodies, washed, and followed by the secondary antibodies. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories, Burlingame, CA) and viewed with an Olympus Fluoview-1000 confocal scanning system under different wave lengths.

Nguyen C., Nguyen J.P., Modi A.P., Ahmad I., Petrova S.C., Ferrell SD J.r., Wilhelm S.R., Ye Y., Schaue D, & Barsky S.H. (2021). Use of constitutive and inducible oncogene-containing iPSCs as surrogates for transgenic mice to study breast oncogenesis. Stem Cell Research & Therapy, 12, 301.

+ Open protocol

+ Expand

Multiparametric Profiling of Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against human TLR4, CD133, Nestin, SSEA-1, Msil, CD28, CD45RA, CD45RO, CCR7, CD44, IFN-γ and isotype control were purchased from eBioscience (San Diego, USA). Antibody against human Nanog was purchased from BD Biosciences (USA). Data were obtained with a flow cytometer (FACS Canto; BD Biosciences, USA).

Che F., Yin J., Quan Y., Xie X., Heng X., Du Y, & Wang L. (2017). TLR4 interaction with LPS in glioma CD133+ cancer stem cells induces cell proliferation, resistance to chemotherapy and evasion from cytotoxic T lymphocyte-induced cytolysis. Oncotarget, 8(32), 53495-53507.

+ Open protocol

+ Expand

Purification and Characterization of PGCLCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

PGCLCs were purified by cell sorting using antibodies against ITGB3 (Biolegend) and SSEA1 (eBioscience) conjugated with PE and Alexa Fluor 647, respectively, as previously described (35 (link),36 ). After dissecting single cells, the cells were resuspended in staining buffer (PBS + 3% FBS) for 20 min on ice, stained with the primary antibodies, washed with staining buffer, resuspended in staining buffer and sorted by FACS. Small debris and doublets were removed by gating for size. For negative controls, we analyzed unstained and secondary antibody only (primary omitted) stained cells. The data were processed using FlowJo software (BD Biosciences). Results were from at least three independent replicates. Statistical significance was determined using the paired Student's t‐test.

Tan K, & Wilkinson M.F. (2022). Regulation of both transcription and RNA turnover contribute to germline specification. Nucleic Acids Research, 50(13), 7310-7325.

+ Open protocol

+ Expand

Characterizing Stem Cell Phenotypes

Check if the same lab product or an alternative is used in the 5 most similar protocols

SSEA-1 (eBioscience) and Sox2 (BD Pharmingen) staining was performed according to the manufacturer's instructions. Flow cytometric analysis was performed with an LSRII flow cytometer (BD Biosciences) and FlowJo software (TreeStar, Ashland, OR). Isotype controls were used in each experiment.

Cheng E.C., Kang D., Wang Z, & Lin H. (2014). PIWI Proteins Are Dispensable for Mouse Somatic Development and Reprogramming of Fibroblasts into Pluripotent Stem Cells. PLoS ONE, 9(9), e97821.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!