Multicolor Immunofluorescent Staining Protocol

The protocol for immunohistochemical staining was as described previously^{30 (link)}. Staining reagents included biotinylated PNA (Vector), AlexaFluor 647 anti-B220, biotinylated anti-IgD, AlexaFluor 488 anti-CD45.2 (Biolegend), and streptavidin AlexaFluor 568 (Invitrogen). All stained slides were mounted with ProlongGold antifade reagents (Invitrogen) and examined with an Olympus FV1000 confocal system.

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Liu X., Chen X., Zhong B., Wang A., Wang X., Chu F., Nurieva R.I., Yan X., Chen P., van der Flier L.G., Nakatsukasa H., Neelapu S.S., Chen W., Clevers H., Tian Q., Qi H., Wei L, & Dong C. (2014). Transcription factor Achaete-Scute homologue 2 initiates T follicular helper cell development. Nature, 507(7493), 513-518.

Publication 2014

biotinylated PNA AlexaFluor 647 anti-B220 biotinylated anti-IgD AlexaFluor 488 anti-CD45.2 streptavidin AlexaFluor 568 ProlongGold antifade reagents FV1000 confocal system

Alexafluor 647 Anti igd Streptavidin Vector

Corresponding Organization :

Other organizations : The University of Texas MD Anderson Cancer Center, Tsinghua University, Wuhan University, Lymphoma Research Foundation, Institute for Systems Biology, National Eye Institute, Utrecht University, National Institutes of Health, Hubrecht Institute for Developmental Biology and Stem Cell Research, University Medical Center Utrecht

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Immunohistochemical staining patterns

control variables

Staining protocol as described previously in reference 30
Staining reagents used (biotinylated PNA, AlexaFluor 647 anti-B220, biotinylated anti-IgD, AlexaFluor 488 anti-CD45.2, and streptavidin AlexaFluor 568)
Microscopy system used (Olympus FV1000 confocal system)
Mounting medium (ProlongGold antifade reagents)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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