STBin TOs were immunostained as described1 (link). For staining of STBout TOs in suspension, the same protocol described was used, but the releasing of organoids from Matrigel was omitted. The following antibodies or reagents were used: SDC-1 (Abcam, ab128936), hCG beta (Abcam, ab243581), ITGA6 (Invitrogen, MA5-16884), HLA-G (Abcam, ab52454 and ab283260), cytokeratin-19 (Abcam, ab9221), Alexa Fluor 488 Goat anti-Mouse IgG secondary antibody (Invitrogen, R37120). Alexa Fluor 488 Goat anti-Rabbit IgG secondary antibody (Invitrogen, R37116), Alexa Fluor 488 Goat anti-Rat IgG secondary antibody (Invitrogen, A11006), Alexa Fluor 594 Goat anti-Mouse IgG secondary antibody (Invitrogen, R37121), Alexa Fluor 594 Goat anti-Rabbit IgG secondary antibody (Invitrogen, R37117), Alexa Fluor 633 Goat anti-Mouse IgG secondary antibody (Invitrogen, A21052), Alexa Fluor 647 Goat anti-Rat IgG secondary antibody (Invitrogen, A21247), Images were captured using a Olympus Fluoview FV3000 inverted confocal microscope or a Zeiss 880 Airyscan Fast Inverted confocal microscope and contrast-adjusted in Photoshop or Fiji. Image analysis and generation of three-dimensional movies was performed using Imaris (version 9.2.1, Oxford Instruments).
Alexa fluor 488 goat anti rat igg secondary antibody
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488 Goat anti-Rat IgG secondary antibody is a fluorescently labeled antibody used for detection and visualization of rat immunoglobulin G (IgG) in various applications, such as immunofluorescence, flow cytometry, and Western blotting.
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Lab products found in correlation
Alexa fluor 594 goat anti mouse igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti mouse igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Imaris, by Oxford Instruments (1 mentions)
2 protocols using alexa fluor 488 goat anti rat igg secondary antibody
1
Immunostaining of Trophoblast Organoids
2
Immunofluorescence Analysis of LC3 II
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The expression of LC3 II was detected using immunofluorescence. In total, ~2×104 cells were seeded into 24-well plates, washed three times with PBS and fixed for 20 min with 4% PFA at room temperature. PBS with 1% Triton X-100 was then added for 20 min. Following three washes with PBS, the cells were blocked for 1 h with 4% goat serum (diluted with PBS) at room temperature. The cells were then incubated with a rabbit monoclonal antibody against LC3 II (1:100; catalog no. ab48394; Abcam) at 4°C overnight. Alexa Fluor® 488 goat anti-rat IgG secondary antibody (1:400; Invitrogen; catalog no. O-6382; Thermo Fisher Scientific, Inc.) was then added for 2 h at room temperature in the dark. DAPI (300 nM) was used for nuclear staining for 15 min at room temperature. Images of ten randomly selected microscopic fields were captured using a confocal microscope (magnification, ×2,000).
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