Cd45.2 fitc clone 104

Manufactured by BD

CD45.2 FITC (clone 104) is a fluorochrome-conjugated antibody that binds to the CD45.2 antigen expressed on the surface of mouse cells. It is commonly used in flow cytometry applications to identify and analyze various leukocyte populations.

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Lab products found in correlation

Prolong antifade, by Thermo Fisher Scientific (1 mentions) Antigenfix, by Microtech (1 mentions)

2 protocols using cd45.2 fitc clone 104

Competitive HSC Transplantation Assay

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C57BL6/J-CD45.1 mice were used as recipients and competitors. The recipient mice were irradiated by 10 Gy divided into 2 fractions on the day –1. Competitor cells were prepared from E14.5 fetal livers from the C57BL6/J-CD45.1 mice and frozen in advance. On day 0, LSK CD150⁺ HSCs were sorted from E14.5 fetal livers of donor embryos that express CD45.2. Each recipient mouse was transplanted with 150 donor HSCs and 3 × 10⁵ competitor fetal liver cells by tail vein injection. After 12 weeks, peripheral blood and bone marrow cells of recipients were analyzed for the presence of CD45.1⁺ and CD45.2⁺ cells by a LSRFortessa. Peripheral blood mononucleic cells were stained with the following antibodies for analyzing myeloid lineage: CD45.1 PE (A20) (BD Biosciences), CD45.2 FITC (clone 104) (BD Biosciences), Mac-1 Ax647 (M1/70) (BD Biosciences), and Gr-1 Pacific Blue (RB6-8C5) (BioLegend). The following antibodies were used for lymphoid lineage: CD45.1 PE (A20), CD45.2 FITC (clone 104), B220 APC (RA3-6B2), and CD3 Pacific Blue (17A2).

Niizuma H., Searleman A.C., Takeda S., Armstrong S.A., Park C.Y., Cheng E.H, & Hsieh J.J. (2021). Taspase1 orchestrates fetal liver hematopoietic stem cell and vertebrae fates by cleaving TFIIA. JCI Insight, 6(15), e149382.

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Multicolor Immunofluorescence Staining Protocol

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Collected spleen fragments were immediately fixed with Antigenfix (Microm Microtech France, Francheville, France) for 3 hr at room temperature, followed by two washes in PBS 0•1 m pH 7•4 and a 30% sucrose bath at 4°, before embedding in OCT. Pieces were kept at -80° and sliced at 8-µm thickness. The mAbs CD45.2 (FITC clone 104; BD Pharmingen), B220 (RA3-6B2 A-647; BD Pharmingen), F4-80 (purified, clone SF12; BD Pharmingen), anti-MHC class II (M5/114-biotin; eBioscience), CD11c (N418 purified; eBioscience) Ki67 (SolA15 purifed; eBioscience) were diluted in PBS, at a pre-established concentration. Slices were incubated overnight at 4°. Secondary labelling was for 2 hr at room temperature. Prolong anti-fade (Invitrogen) containing or not DAPI was used to keep the coloured slices. Confocal microscopy was performed with the Zeiss 780 microscope with a 40× oil objective and a 0•6× zoom.

Mouchacca P., Chasson L., Frick M., Foray C., Schmitt-Verhulst A.M, & Boyer C. (2015). Visualization of granzyme B-expressing CD8 T cells during primary and secondary immune responses to Listeria monocytogenes. Immunology, 145(1).

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