Cells were fixed in BD CytofixTM fixation buffer (BD Biosciences) for 15 min at room temperature and then washed three times with BD perm/wash buffer (BD Biosciences) every 10 min. After a 30 min incubation with perm/wash buffer at room temperature, cells were stained with the following conjugated primary antibodies and incubated overnight at 4°C: OCT3/4-FITC (1:100; BD Pharmingen), NANOG-PE (1:100; BD Pharmingen), SOX2-PE (1:50; BD Biosciences), SSEA3-PE (1:100; BD Pharmingen), SSEA4-FITC (1:100; BD Biosciences), TRA-1-60-FITC (1:100; BD Pharmingen), and βIII-tubulin-FITC (1:100; BD Biosciences). Cells were imaged using fluorescence microscopy (Leica, Wetzlar, Germany).
Ssea4 fitc
Manufactured by BD
SSEA4-FITC is a fluorescent-conjugated antibody that detects the stage-specific embryonic antigen 4 (SSEA4), a carbohydrate marker expressed on the surface of undifferentiated human embryonic stem cells and certain types of cancer cells. This product is designed for use in flow cytometry and other cell-based applications to identify and characterize SSEA4-positive cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Cd90 apc, by BD (2 mentions)
Cd44 apc, by BD (2 mentions)
Cd45 allophycoerythrin apc, by BD (2 mentions)
Facscanto 2, by BD (2 mentions)
Cd33 bv421, by BioLegend (2 mentions)
Cd34 pe cy7, by BioLegend (2 mentions)
Facscanto 2 flow cytometer, by BD (2 mentions)
Perm wash buffer, by BD (2 mentions)
Cytofix fixation buffer, by BD (1 mentions)
Nanog pe, by BD (1 mentions)
Ssea3 pe, by BD (1 mentions)
Fluorescence microscopy, by Leica (1 mentions)
Sox2 pe, by BD (1 mentions)
Cd31 phycoerythrin pe, by BD (1 mentions)
Cd146 fitc, by BD (1 mentions)
Cd34 fluorescein isothiocyanate fitc, by BD (1 mentions)
5 protocols using ssea4 fitc
1
Comprehensive Stem Cell Characterization
2
Immunophenotyping of PVC from Normal and GDM Pregnancies
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Single cell suspensions were prepared from normal pregnant women and GDM patient-derived HUCPVC (hereafter called N-PVC and GDM-PVC) cultures by dissociation with 0.05% trypsin-EDTA. The cells were resuspended in 1% FBS-PBS and were stained for 1 hr with the following fluorochrome-conjugated anti-human antibodies: CD31-phycoerythrin (PE), CD34-luorescein-isothiocyanate (FITC), CD45-allophycoerythrin (APC), SSEA-4-FITC, CD146-FTIC, CD44-APC and CD90-APC (all BD Biosciences) or their corresponding isotype controls. After immunostaining, the cells were incubated with the viability dye 7-aminoactinomycin D (7AAD) to exclude dead cells. Data was analyzed using FACSCanto II (BD Biosciences) and FlowJo software.
3
Pluripotency and Hematopoietic Differentiation Assays
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To assess the pluripotency of iPS cells, the antibodies TRA1-60-PE (Cat Nr. MA1-023-PE, eBioscience) and SSEA4-FITC (Cat Nr. 560126, BD biosciences, BD) were used. Dead cells were excluded from the analysis by 4′,6-diamidino-2-phenylindole (DAPI; 1μg/ml) (Cat Nr. D3571, Thermo Fisher Scientific) staining. For detection of hematopoietic progenitor cells, the antibodies CD33-BV421 (Cat Nr. 366622, BioLegend, BL), CD34-PE-Cy7 (Cat Nr. 343615, BL), KDR-AF647 (Cat Nr. 359909, BL), CD43-PE (Cat Nr. 343204, BL), CD41a-FITC (Cat Nr. 303703, BL), CD235a-FITC (Cat Nr. 349103, BL), CD45-BV510 (Cat Nr. 103138, BL) and 7-AAD (Cat Nr. 420404, BL) were used as an “early-stage” multicolor hematopoietic cell panel. For the detection of mature myeloid cells, the antibodies CD15-PE (Cat Nr. 301905, BL), CD16-FITC (Cat Nr. 302005, BL), CD14-APC-H7 (Cat Nr. 367117, BL), CD45-BV510 (Cat Nr. 103138, BL), CD33 BV-421 (Cat Nr. 366622, BL) and 7-AAD (Cat Nr. 420404, BL) were used as a “late-stage” multicolor myeloid differentiation panel. Anti-mouse IgGk beads were used for compensation. Antibodies and beads for flow cytometry were purchased from BD Biosciences unless otherwise indicated. Samples were analyzed using a FACS Canto II flow cytometer (Becton-Dickinson) and FlowJo software (FLOWJO, LLC, Ashland, OR).
4
Characterization of Human Umbilical Cord Perivascular Cells
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HUCPVCs were dissociated into single cells using trypsin-EDTA and resuspended in
1% FBS-PBS. The cells were filtered through a 70 µm cell strainer and reacted
with the following antibodies conjugated with fluorochrome for 1 hr at 4℃:
CD31-phycoerythrin (PE), CD34-fluorescein-isothiocyanate (FITC),
CD45-allophycoerythrin (APC), CD44-APC, CD90-APC, CD146-FITC, and SSEA-4-FITC
(all, BD Biosciences). The dead cells were excluded with 7-aminoactinomycin D.
Stained cells were analyzed using the FACSCanto II (BD Biosciences) and data
were analyzed by FlowJo software (FlowJo).
1% FBS-PBS. The cells were filtered through a 70 µm cell strainer and reacted
with the following antibodies conjugated with fluorochrome for 1 hr at 4℃:
CD31-phycoerythrin (PE), CD34-fluorescein-isothiocyanate (FITC),
CD45-allophycoerythrin (APC), CD44-APC, CD90-APC, CD146-FITC, and SSEA-4-FITC
(all, BD Biosciences). The dead cells were excluded with 7-aminoactinomycin D.
Stained cells were analyzed using the FACSCanto II (BD Biosciences) and data
were analyzed by FlowJo software (FlowJo).
5
Pluripotency and Hematopoietic Differentiation Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the pluripotency of iPS cells, the antibodies TRA1-60-PE (Cat Nr. MA1-023-PE, eBioscience) and SSEA4-FITC (Cat Nr. 560126, BD biosciences, BD) were used. Dead cells were excluded from the analysis by 4′,6-diamidino-2-phenylindole (DAPI; 1μg/ml) (Cat Nr. D3571, Thermo Fisher Scientific) staining. For detection of hematopoietic progenitor cells, the antibodies CD33-BV421 (Cat Nr. 366622, BioLegend, BL), CD34-PE-Cy7 (Cat Nr. 343615, BL), KDR-AF647 (Cat Nr. 359909, BL), CD43-PE (Cat Nr. 343204, BL), CD41a-FITC (Cat Nr. 303703, BL), CD235a-FITC (Cat Nr. 349103, BL), CD45-BV510 (Cat Nr. 103138, BL) and 7-AAD (Cat Nr. 420404, BL) were used as an “early-stage” multicolor hematopoietic cell panel. For the detection of mature myeloid cells, the antibodies CD15-PE (Cat Nr. 301905, BL), CD16-FITC (Cat Nr. 302005, BL), CD14-APC-H7 (Cat Nr. 367117, BL), CD45-BV510 (Cat Nr. 103138, BL), CD33 BV-421 (Cat Nr. 366622, BL) and 7-AAD (Cat Nr. 420404, BL) were used as a “late-stage” multicolor myeloid differentiation panel. Anti-mouse IgGk beads were used for compensation. Antibodies and beads for flow cytometry were purchased from BD Biosciences unless otherwise indicated. Samples were analyzed using a FACS Canto II flow cytometer (Becton-Dickinson) and FlowJo software (FLOWJO, LLC, Ashland, OR).
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