For immunostaining, cells were cultured in matrigel-coated Lab-Tek Chamber slides (Nunclone), fixed in 4% paraformaldehyde in PBS, permeabilized with 0.5% Triton X-100 and blocked in PBS supplemented with 5% FBS. Cells were stained by specific antibodies; SSEA-4 and TRA-1–60 (Millipore #SCR001), OCT4 (Cell Signaling Technology #2750), SOX2 (Cell Signaling Technology #2748), FOXA2 (Millipore #07-633), β-III tubulin (Abcam # 41489), CD31 (Santa Cruz Biotechnology # SC1506), cardiac troponin T (Abcam #ab8295).
Foxa2
Manufactured by Merck Group
Sourced in United States
FOXA2 is a protein-coding gene that produces the forkhead box protein A2, a transcription factor involved in regulating gene expression. It plays a crucial role in embryonic development and the maintenance of mature tissues. The FOXA2 protein is essential for the proper formation and function of various organ systems.
Automatically generated - may contain errors
Lab products found in correlation
βiii tubulin, by Abcam (3 mentions)
Muc5b, by Santa Cruz Biotechnology (1 mentions)
Muc5ac, by Santa Cruz Biotechnology (1 mentions)
C peptide, by Cell Signaling Technology (1 mentions)
Lsm 780, by Zeiss (1 mentions)
Insulin, by Merck Group (1 mentions)
Tra 1 60, by Merck Group (1 mentions)
Nanog, by Abcam (1 mentions)
Sox17, by R&D Systems (1 mentions)
Hnf4a, by Santa Cruz Biotechnology (1 mentions)
Dabco, by Merck Group (1 mentions)
Olig2, by Merck Group (1 mentions)
Stemdiff trilineage differentiation kit, by STEMCELL (1 mentions)
Af1924, by R&D Systems (1 mentions)
Af2085, by R&D Systems (1 mentions)
Mab5326, by Merck Group (1 mentions)
Ab5790, by Abcam (1 mentions)
7 protocols using foxa2
1
Immunostaining Pluripotency and Lineage Markers
2
Quantifying Nasal Epithelial Cell Types
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Human nasal tissues were stained with PAS, and the PAS-positive cells, which displayed a purple color, were counted via light microscopy (400×) and expressed as the percentage of the total epithelial cells (500 cells counted). IHC was performed as reported elsewhere,13 (link) and the sections were stained using mouse monoclonal antibodies against the following proteins: FoxA2 (1:50; Millipore, Billerica, MA, USA), MUC5AC, and MUC5B (1:100; Santa Cruz Biotech, Santa Cruz, CA, USA). Then, the antibodies were detected via streptavidin-biotin-horseradish peroxidase complex formation. The immunostaining result was considered positive when brown cells appeared following reaction with the reagent 3', 3'-diaminobenzidine. Replacement of primary antibodies with isotype-matched IgG was used as a negative control. The sections were examined via light microscopy (400×), and the patterns of antibody staining were scored in a quantitative manner. The pattern of immunoreactivity was analyzed in the epithelium and subepithelial area. The number of immunoreactive cells (stained brown) was expressed as the percentage of the total cell number (500 cells counted). The immunoreactivity score was assessed by 2 independent observers in a blinded manner, and their results were averaged.
3
Immunostaining of iPSCs and Tissue Samples
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For immunostaining of iPSCs and iPSC progeny, cells were cultured in Matrigel-coated Lab-Tek Chamber slides (Nunclone). For immunostaining of pancreatic or renal samples, tissues were embedded and frozen in OCT compound (Sakura, Torrance, CA, http://www.sakura-americas.com ). Cells/tissues were then fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and blocked in PBS with 5% fetal bovine serum (FBS). Cells were then stained with specific antibodies, including insulin (catalog no. 12018, Sigma-Aldrich, St. Louis, MO, http://www.sigmaaldrich.com ), C-peptide (catalog no. 4593; Cell Signaling Technology, Beverly, MA, http://www.cellsignal.com ), TRA-1-60 (catalog no. SCR001; EMD Millipore, Billerica, MA, http://www.millipore.com ), OCT4 (catalog no. 2750; Cell Signaling Technology), SOX2 (catalog no. 2748; Cell Signaling Technology), NANOG (catalog no. ab21624; Abcam, Cambridge, MA, http://www.abcam.com/ ), FOXA2 (catalog no. 07-633; Millipore), β-III tubulin (catalog no. 41489; Abcam), and CD31 (catalog no. SC1506; Santa Cruz Biotechnology, Santa Cruz, CA, http://www.scbt.com ). Nuclei were counterstained by 4′,6-diamidino-2-phenylindole (DAPI). Stained cells were observed through a Zeiss LSM 780 (Zeiss, Stuttgart, Germany, http://www.zeiss.com ) confocal laser scanning microscope, and the images were analyzed by using the Zeiss imaging software.
4
Immunofluorescence Staining of Hepatic Markers
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Cells were fixed with 4% paraformaldehyde for 15 min and washed twice with phosphate-buffered saline. The cell membrane was permeabilized using 0.1% Triton X-100 in phosphate-buffered saline for 10 min. After blocking the cells with 5% FBS in phosphate-buffered saline, the cells were incubated with one or two of the following primary antibodies followed by a secondary antibody: FOXA2 (07-633, Millipore), SOX17 (AF1924, R&D Systems), HNF4A (sc-6556, Santa Cruz Biotechnology), TBX3 (ab99302, Abcam), CPS1 (Hepatocytes Monoclonal Antibody (OCH1E5); MA5-12417, Thermo Fisher Scientific), AFP (A8462, Sigma), and albumin (A6684, Sigma).
5
Immunofluorescence Analysis of Mouse Embryos
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Mouse embryos were fixed in 4% paraformaldehyde (PFA) in phosphate-balanced saline (PBS) for up to 1 h, washed with PBS briefly and left in 30% sucrose overnight, embedded in O.C.T freezing media and frozen at −80 °C. Cryosections at 10 µm thickness were cut with a Leica LM1900 Cryostat. For immunofluorescence assays, sections were allowed to dry at room temperature for 1 h, blocked in blocking buffer (PBS plus 0.1% Triton X-100 and 1% goat serum). They were then incubated in blocking buffer with appropriate primary antibodies at 4 °C overnight, washed in blocking buffer three times and incubated in blocking buffer with Cy3-conjugated secondary antibodies, wash three more times and mounted with DABCO (Sigma-Aldrich, Saint Louis, MO, USA). Antibodies used: Foxa2, Nk2.2, Nkx6.1, Pax6, Pax7, Shh (DSHB) and Olig2 (Millipore, AB9610). Photos were taken on a Nikon E600 microscope with a Micropublisher CCD camera (QImaging, Surrey, BC, Canada).
6
Immunostaining Pluripotency and Lineage Markers
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For immunostaining, cells were cultured in matrigel-coated Lab-Tek Chamber slides (Nunclone), fixed in 4% paraformaldehyde in PBS, permeabilized with 0.5% Triton X-100 and blocked in PBS supplemented with 5% FBS. Cells were stained by specific antibodies; SSEA-4 and TRA-1–60 (Millipore #SCR001), OCT4 (Cell Signaling Technology #2750), SOX2 (Cell Signaling Technology #2748), FOXA2 (Millipore #07-633), β-III tubulin (Abcam # 41489), CD31 (Santa Cruz Biotechnology # SC1506), cardiac troponin T (Abcam #ab8295).
7
Three-Germ Layer Differentiation Assay
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Alternative three-germ layer derived cells were differentiated using a commercially available kit, STEMdiff Trilineage Differentiation Kit (Stem Cell Technologies). Cells were derived according to manufacturer’s instructions. Gene expression was evaluated using qPCR. Immunostaining was performed using a FOXA2 (07-633, Millipore) and a SOX17 (AF1924, R&D) for the endoderm, T (AF2085, R&D) and NCAM (362502, BioLegend) for the mesoderm, PAX6 (ab5790, abcam), and a NESTIN (MAB5326, Millipore) for the ectoderm.
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