Protocol detail

Multicolor Immunofluorescence Staining Protocol

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These procedures were carried out as described previously (22 (link), 23 (link)). Antibodies used for staining: purified rat anti-mouse CD35 (8c12; BD Biosciences), biotin-anti-CD45.2 (104; BD Biosciences), Alexa Fluor 555-conjugated goat polyclonal anti-rat (Invitrogen), Alexa Fluor 488-Streptavidin (Invitrogen), Alexa Fluor 647-conjugated rat Ab to mouse IgD (11–26C.2a; Biolegend). Mounted sections were imaged with 10 × and 20 × objective on a Nikon Eclipse Ti-S microscope.

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Geng J., Wei H., Shi B., Wang Y.H., Greer B.D., Pittman M., Smith E., Thomas P.G., Kutsch O, & Hu H. (2019). Bach2 negatively regulates Tfh cell differentiation and is critical for CD4+ T cell memory. Journal of immunology (Baltimore, Md. : 1950), 202(10), 2991-2998.

Publication 2019

rat anti-mouse CD35 (8c12; biotin-anti-CD45.2 (104; Alexa Fluor 555-conjugated goat polyclonal anti-rat Alexa Fluor 488-Streptavidin Eclipse Ti-S microscope

Alexa fluor 488 Alexa fluor 555 Alexa fluor 647 Antibodies Biotin 2 Cd35 Goat Microscope Mouse Streptavidin

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Variable analysis

independent variables

Purified rat anti-mouse CD35 (8c12; BD Biosciences)
Biotin-anti-CD45.2 (104; BD Biosciences)
Alexa Fluor 555-conjugated goat polyclonal anti-rat (Invitrogen)
Alexa Fluor 488-Streptavidin (Invitrogen)
Alexa Fluor 647-conjugated rat Ab to mouse IgD (11–26C.2a; Biolegend)

dependent variables

Stained cells imaged with 10 × and 20 × objective on a Nikon Eclipse Ti-S microscope

control variables

Procedures carried out as described previously (22, 23)

controls

Positive controls not specified
Negative controls not specified

Annotations

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