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Mouse anti ssea4

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Mouse anti-SSEA4 is a monoclonal antibody that recognizes the SSEA-4 (Stage-Specific Embryonic Antigen 4) surface marker. SSEA-4 is a glycosphingolipid that serves as a marker for undifferentiated embryonic stem cells and other pluripotent cells.

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Lab products found in correlation

Mouse anti tra 1 60, by Cell Signaling Technology (3 mentions) Ab13970, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Mouse anti nestin, by Abcam (2 mentions) Rabbit anti tom20, by Santa Cruz Biotechnology (2 mentions) Mouse anti tra 1 81, by Cell Signaling Technology (2 mentions) Alexa fluor 555, by Thermo Fisher Scientific (2 mentions) Alexa fluor 647, by Thermo Fisher Scientific (2 mentions) Z1 axio observer apotome, by Zeiss (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Rabbit anti pax6, by Fortrea (2 mentions) Goat anti sox2, by Santa Cruz Biotechnology (2 mentions) Goat anti oct3 4, by Santa Cruz Biotechnology (2 mentions) Mouse anti tra 1 81, by Merck Group (2 mentions) Tcs sp5 laser scanning microscope, by Leica (2 mentions) Rabbit anti foxa2, by Cell Signaling Technology (2 mentions) Goat anti pdx1, by R&D Systems (1 mentions) Mouse anti glucagon, by Merck Group (1 mentions) Rabbit anti oct4, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Mouse anti-human SSEA4 Anti-SSEA4 SSEA4

5 protocols using mouse anti ssea4

1

Immunocytochemistry and Mitochondrial Analysis

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Cells were grown in coated 96-well plates or chamber slides, fixed in 4% paraformaldehyde for 10 min, and permeabilized in 0.3% Triton X-100 in PBS for 10 min. Cells were then blocked with 5% BSA for at least 1 hr before incubation with chicken anti-GFP (Abcam, ab13970) 1:2000, rabbit anti-Tom20 (Santa Cruz, SC11415) 1:500, mouse anti-SSEA4 (Cell Signaling CS 4755) 1:500, mouse anti Tra-1-60 (Cell Signaling CS 4746) 1:500, mouse anti Tra-1-81 (Cell Signaling CS 4745) 1:500, rabbit anti-Pax6 (Covance, PRB-278P) 1:500, or mouse anti-Nestin (Abcam, ab6142) 1:1000 overnight at 4°C. Cells were washed 3 times with PBS, then incubated with AlexaFluor 488, AlexaFluor 555, and/or AlexaFluor 647 (Fisher) at 1:500 dilutions for 1 hr at RT. To stain nuclei, cells were incubated with 1:10000 Hoechst 33342 (Life Technologies). Cells were preserved in Prolong Gold anti-fade reagent (Life Technologies) and imaged on a fluorescence microscope (Z1 Axio Observer Apotome, Zeiss). Mitochondrial length was quantified using an ImageJ mitochondrial morphometry plugin as described previously (Dickey and Strack, 2011 (link)).
Ward J.M., Stoyas C.A., Switonski P.M., Ichou F., Fan W., Collins B., Wall C.E., Adanyeguh I., Niu C., Sopher B.L., Kinoshita C., Morrison R.S., Durr A., Muotri A.R., Evans R.M., Mochel F, & La Spada A.R. (2019). Metabolic and Organelle Morphology Defects in Mice and Human Patients Define Spinocerebellar Ataxia Type 7 as a Mitochondrial Disease. Cell reports, 26(5), 1189-1202.e6.
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2

Immunofluorescence Analysis of Stem Cell Markers

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Cells were fixed with 4% paraformaldehyde for 30 min and then permeabilized in PBS containing 0.2% Triton X-100. Cells were blocked with PBS containing 3% BSA, and incubated with primary antibodies overnight at 4 °C. Then the cells were incubated with the secondary antibodies for 1 h at room temperature after washing with PBS. Images were acquired on a TCS SP5 laser-scanning microscope (Leica). The following antibodies and dilutions were used: goat anti-OCT-3/4 (1:500, Cat #sc-8628, Santa Cruz), goat anti-SOX2 (1:500, Cat #sc-17320, Santa Cruz), mouse anti-TRA-1-81 (1:50, Cat #MAB4381, Millipore), mouse anti-SSEA4 (1:500, Cat #4755, Cell Signaling), rabbit anti-FOXA2 (1:250, Cat #8186, Cell Signaling), goat anti-SOX17 (1:500, Cat #GT15094, Acris/Novus), goat anti-PDX1 (1:500, Cat #AF2419, R&D Systems), rabbit anti-NKX6.1 (1:300, Cat #NBP1-82553, Acris/Novus), guinea pig anti-C-Peptide (1:100, Cat #ab30477, Abcam), mouse anti-Glucagon (1:500, Cat #G2654, Sigma), rat anti-Somatostatin (1:300, Cat #MA5-16987, Invitrogen).
Wang X., Sterr M., A.n.s.a.r.u.l.l.a.h., Burtscher I., Böttcher A., Beckenbauer J., Siehler J., Meitinger T., Häring H.U., Staiger H., Cernilogar F.M., Schotta G., Irmler M., Beckers J., Wright C.V., Bakhti M, & Lickert H. (2019). Point mutations in the PDX1 transactivation domain impair human β-cell development and function. Molecular Metabolism, 24, 80-97.
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3

Immunofluorescence Staining of Stem Cells

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Immunofluorescence was performed on cells grown directly on slides as previously described (Chadwick and Willard 2002 (link)). Except, the maximum slide seed area was delimited using a Super PAP Pen (IM3580, Beckman Coulter®) and coated with 700µl of Matrigel™ or 0.1% Gelatin prior to seeding hESCs or EBOGs, respectively. Additionally, hESCs were allowed to adhere for 1hr in complete StemPro® media supplemented with ROCK Inhibitor Y-27632 (ROCKi, SCM075, Millipore™), the media was then replaced with fresh complete StemPro® media (-ROCKi), and allowed to recover overnight. Antibodies used for indirect Immunofluorescence included rabbit anti-H3K4me2 (07–030, EMD Millipore), mouse anti-H3K27me3 (ab6002, abcam), rabbit anti-NANOG (3580S, Cell Signaling Technology), mouse anti-SSEA4 (4755S, Cell Signaling Technology), goat anti-SOX2 (AF2018, R&D Systems), and rabbit anti-OCT4 (2750S, Cell Signaling Technology). Conjugate secondary antibodies (Alexa-Fluor®) were obtained from Life Technologies Corporation. DNA was counterstained using the VECTASHIELD® mounting medium with DAPI from VECTOR Laboratories. Imaging was performed on an Olympus IX71 operated by the DeltaVision pDV, deconvolved with softWoRx 5.5.1 (DeltaVision), and compiled using Adobe Photoshop CS6 (Adobe Systems).
Figueroa D.M., Darrow E.M, & Chadwick B.P. (2015). Two novel DXZ4-associated long noncoding RNAs show developmental changes in expression coincident with heterochromatin formation at the human (Homo sapiens) macrosatellite repeat. Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology, 23(4), 733-752.
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4

Immunofluorescence Characterization of Stem Cells

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Cells were fixed with 4% paraformaldehyde for 30 min and then permeabilized in PBS containing 0.2% Triton X-100. Cells were blocked with PBS containing 3% BSA, and incubated with primary antibodies overnight at 4 °C. Then secondary antibodies were incubated for 1 h at room temperature after washing with PBS. Images were acquired on a TCS SP5 laser-scanning microscope (Leica). The following antibodies and dilutions were used: goat anti-OCT-3/4 (1:500, #sc-8628, Santa Cruz), goat anti-SOX2 (1:500, #sc-17320, Santa Cruz), mouse anti-TRA-1-60 (1:1000, #4746, Cell Signaling), mouse anti-TRA-1-81 (1:50, MAB4381, Millipore), mouse anti-SSEA4 (1:500, #4755, Cell Signaling), rabbit anti-FOXA2 (1:250, #8186, Cell Signaling), goat anti-SOX17 antibody (1:500, #GT15094, Acris/Novus), goat anti-PDX1 antibody (1:500, #AF2419, R&D Systems).
Wang X., Sterr M., Burtscher I., Chen S., Hieronimus A., Machicao F., Staiger H., Häring H.U., Lederer G., Meitinger T., Cernilogar F.M., Schotta G., Irmler M., Beckers J., Hrabě de Angelis M., Ray M., Wright C.V., Bakhti M, & Lickert H. (2018). Genome-wide analysis of PDX1 target genes in human pancreatic progenitors. Molecular Metabolism, 9, 57-68.
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5

Immunocytochemistry and Mitochondrial Analysis

Cited 1 time
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Cells were grown in coated 96-well plates or chamber slides, fixed in 4% paraformaldehyde for 10 min, and permeabilized in 0.3% Triton X-100 in PBS for 10 min. Cells were then blocked with 5% BSA for at least 1 hr before incubation with chicken anti-GFP (Abcam, ab13970) 1:2000, rabbit anti-Tom20 (Santa Cruz, SC11415) 1:500, mouse anti-SSEA4 (Cell Signaling CS 4755) 1:500, mouse anti Tra-1-60 (Cell Signaling CS 4746) 1:500, mouse anti Tra-1-81 (Cell Signaling CS 4745) 1:500, rabbit anti-Pax6 (Covance, PRB-278P) 1:500, or mouse anti-Nestin (Abcam, ab6142) 1:1000 overnight at 4°C. Cells were washed 3 times with PBS, then incubated with AlexaFluor 488, AlexaFluor 555, and/or AlexaFluor 647 (Fisher) at 1:500 dilutions for 1 hr at RT. To stain nuclei, cells were incubated with 1:10000 Hoechst 33342 (Life Technologies). Cells were preserved in Prolong Gold anti-fade reagent (Life Technologies) and imaged on a fluorescence microscope (Z1 Axio Observer Apotome, Zeiss). Mitochondrial length was quantified using an ImageJ mitochondrial morphometry plugin as described previously (Dickey and Strack, 2011 (link)).
Ward J.M., Stoyas C.A., Switonski P.M., Ichou F., Fan W., Collins B., Wall C.E., Adanyeguh I., Niu C., Sopher B.L., Kinoshita C., Morrison R.S., Durr A., Muotri A.R., Evans R.M., Mochel F, & La Spada A.R. (2019). Metabolic and Organelle Morphology Defects in Mice and Human Patients Define Spinocerebellar Ataxia Type 7 as a Mitochondrial Disease. Cell reports, 26(5), 1189-1202.e6.
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