3D Sw71 models or 2D Sw71 monolayers were cultured for 24–48 h in sterile culture chambers on a glass slide (Falcon). Trophoblast derived explants were cultured in the same type of chambers for expansion of EVTs, and after that the explants were removed carefully. The cells were fixed in 2% paraformaldehyde (PFA) in PBS, overnight (ON) at RT and stained for HLA-C using indirect immunofluorescent method. After washing with PBS, the cells were incubated with 10% goat serum (to block the non-specific binding) and then subsequently with purified polyclonal rabbit anti-human HLA-C antibody (E-AB-17922, Elabscience) and AF488-conjugated goat anti-rabbit antibody (E-AB-1055, Elabscience). Negative controls were prepared by omitting primary antibody and/or secondary antibody. Control staining was done with mouse polyclonal anti-human HLA-G antibody (E-AB-18031, Elabscience) and goat anti-mouse IgG (E-AB-1015, Elabscience). The slides were imaged with ECHO Revolve microscope (RVL-100-M, Echo, San Diego, CA, USA).
E ab 18031
Manufactured by Elabscience
The E-AB-18031 is a laboratory equipment designed for DNA extraction and purification. It utilizes a silica-based membrane technology to efficiently isolate and purify DNA samples from various sources. The core function of this product is to enable the extraction and purification of DNA for further analysis and downstream applications.
Automatically generated - may contain errors
2 protocols using e ab 18031
1
Immunofluorescent Staining of HLA-C
2
Immunofluorescence Analysis of HLA-G and HLA-C in 2D and 3D Sw71 Cell Models
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Sw71 and Sw71-Tw-KO cells as monolayer and spheroids were used in this study. The characterization of these cells has been reported in numerous publications (49) (50) (51) . 3D Sw71 models or 2D Sw71 monolayers were cultured for 24-48 hrs in sterile culture chambers on a glass slide (Falcon). The cells were fixed in 2% paraformaldehyde (PFA) in PBS, overnight (ON) at RT and stained for HLA-G and HLA-C using indirect immunofluorescent method. After washing with PBS, the cells were incubated with Super Block (SkyTec Laboratories, Logan, UT, USA) to suppress the non-specific binding. As primary antibodies we used anti-human rabbit polyclonal antibodies against HLA-G (E-AB-18031, Elabscience) and HLA-C (E-AB-17922, Elabscience) in appropriate dilutions for overnight incubation at 4°C. As a secondary antibody goat anti-rabbit IgG conjugated with AF488 (E-AB-1055, Elabscience) was applied. Negative controls were prepared by omitting primary antibody and/or secondary antibody.
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