OCT-embedded frozen (8 µm), or formalin-fixed and paraffin-embedded (5 µm) tissue sections were used for histology and immunostaining. Immunofluorescence staining and immunohistochemistry were carried out with antibodies described below. For formalin-fixed tissues, antigen retrieval was performed with citrate buffer at 92 °C for 30 min. Antibodies against Ki-67 (#14-5698, 1:300) were obtained from eBioscience, ph-H3 (#3377, 1:400) from Cell Signaling, Cox-2 (#160106, 1:200) from Cayman Chemical, Krt7 (#ab181598, 1:600) from Abcam, and Krt5 (#PRB-160P and # SIG-3475, 1:600), Krt6 (#PRB-169P, 1:600), Krt15 (#PCK-153P, 1:400), CD324 (#147306, 1:300), CD45 (#103101, 1:400), Gr-1 (#108417, 1:400), Ly-6G (#127625, 1:400), F4/80 (#123101, 1:600), and CD3 (#100201, 1:400) from BioLegend.
Cd324
Manufactured by BioLegend
Sourced in United States
CD324, also known as E-cadherin, is a cell adhesion molecule that plays a critical role in the formation and maintenance of cell-cell junctions. It is expressed on the surface of epithelial cells and is essential for the regulation of cell-cell adhesion, cell polarity, and tissue integrity. CD324 is a key marker for the identification and characterization of epithelial cells and is widely used in various research applications.
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Lab products found in correlation
Cox 2, by Cayman Chemical (1 mentions)
F4 80, by BioLegend (1 mentions)
Ssea4, by BioLegend (1 mentions)
Tra 1 60, by Cell Signaling Technology (1 mentions)
Cd146, by BioLegend (1 mentions)
Dmi3000 microscope, by Leica (1 mentions)
Fc500 flow cytometer, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions)
2 protocols using cd324
1
Immunostaining of Tissue Sections
2
Surface Marker Characterization of Human Amniotic Epithelial Cells
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Flow cytometry was used to identify the surface marker expression of the hAECs. After incubation with primary antibodies, including antibodies against HLADR (1:1000, BioLegend, San Diego, USA), CD146 (mesenchymal marker, 1:1000, BioLegend), CD34 (hematopoietic marker, 1:1000, BioLegend), CD324 (epithelial marker, 1:1000, BioLegend), and SSEA4 (pluripotent marker, 1:1000, BioLegend), the hAECs were washed and resuspended in staining buffer. Flow cytometry was immediately performed using an FC500 flow cytometer (BD Pharmingen, San Diego, CA, USA).
For further characterizing the hAECs, immunofluorescence staining was performed. Paraformaldehyde-fixed cells were blocked with 10% goat serum. Then, the cells were incubated with the following primary antibodies: cytokeratin 18 (CK18, an epithelial marker, 1:200, Boster Biological Technology, Wuhan, China) and TRA-1-60 (stem cell marker, 1:1000, Cell Signaling Technology, CST, Beverly, MA, USA). After washing, the hAECs were incubated with secondary antibodies conjugated to Alexa Fluor 488 and 594 (1:3000, CST) and counterstained with DAPI (Abcam, Cambridge, UK). Finally, the immunofluorescence signals were observed with a Leica DMI3000 microscope (Heidelberg, Germany). The negative control cells received identical treatments, except primary antibodies were omitted, and exhibited no specific staining.
For further characterizing the hAECs, immunofluorescence staining was performed. Paraformaldehyde-fixed cells were blocked with 10% goat serum. Then, the cells were incubated with the following primary antibodies: cytokeratin 18 (CK18, an epithelial marker, 1:200, Boster Biological Technology, Wuhan, China) and TRA-1-60 (stem cell marker, 1:1000, Cell Signaling Technology, CST, Beverly, MA, USA). After washing, the hAECs were incubated with secondary antibodies conjugated to Alexa Fluor 488 and 594 (1:3000, CST) and counterstained with DAPI (Abcam, Cambridge, UK). Finally, the immunofluorescence signals were observed with a Leica DMI3000 microscope (Heidelberg, Germany). The negative control cells received identical treatments, except primary antibodies were omitted, and exhibited no specific staining.
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