Cd90.2 clone 30 h12

Manufactured by BioLegend

CD90.2 (clone 30-H12) is a cell surface marker that is expressed on various cell types, including hematopoietic stem cells, neural stem cells, and mesenchymal stem cells. It is commonly used in flow cytometry and cell sorting applications to identify and isolate these cell populations.

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CD90.2 (30-H12), (4 citations)

4 protocols using cd90.2 clone 30 h12

Multiparameter Flow Cytometry Panel

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CD3 clone 17A2 (BioLegend 100237 and 100244), CD4 clone RM4–5 (BioLegend 100545 and 100510), CD4 clone GK13 (BioLegend 100403), CD8 clone 53–6.7 (BioLegend 100734), CD11b clone M1/70 (BioLegend 101257), CD11c clone N418 (BioLegend 117339 and 117338), CD19 clone 6D5 (BioLegend 115522), CD24 clone M1/69 (BioLegend 101822), CD45 clone 30-F11 (BioLegend 103139, 103132, and 103114; eBioscience 56–0451-82), CD45R clone RA3–6B2 (BioLegend 103247, 103246, and 103226), CD69 clone H1.2F3 (eBioscience 25–0691-81), CD90.2 clone 30-H12 (BioLegend 105331), CD103 clone 2E7 (BioLegend 121406 and 121414), F4/80 clone BM8 (BioLegend 123108), Flt3L (R&D Systems AF427), Ly6C clone HK1.4 (BioLegend 128037), Ly6G clone 1A8 (BioLegend 127645), MHC-II clone M5/114.15.2 (BioLegend 707631), NK1.1 clone PK136 (BioLegend 108707, 108720, and 108749), Streptavidin-Brilliant Violet 650 (BioLegend 405231), Streptavidin-APC (eBioscience 17–4317-82). Depleting antibodies: NK1.1 clone PK136 (BioXCell BE0036), and IgG2a isotype control (BioXCell BE0085).

Barry K.C., Hsu J., Broz M.L., Cueto F.J., Binnewies M., Combes A.J., Nelson A.E., Loo K., Kumar R., Rosenblum M.D., Alvarado M.D., Wolf D.M., Bogunovic D., Bhardwaj N., Daud A.I., Ha P.K., Ryan W.R., Pollack J.L., Samad B., Asthana S., Chan V, & Krummel M.F. (2018). A Natural Killer/Dendritic Cell Axis Defines Responsive Tumor Microenvironments in Melanoma. Nature medicine, 24(8), 1178-1191.

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Flow Cytometry Analysis of Transplanted Lungs

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Single cell suspensions were prepared from transplanted lungs as previously described (10 (link)). Cells were stained with fluorochrome-labeled antibodies against CD90.2 (clone 30-H12, Biolegend, San Diego, CA), CD8 (clone 53–6.7, Thermo Fisher Scientific, Waltham MA), CD4 (clone RM4–5, Thermo Fisher Scientific), CD45 (clone 30-F11, Thermo Fisher Scientific), Ki67 (clone SolA15, Thermo Fisher Scientific), CD11b (clone M1/70, Thermo Fisher Scientific), CD11c (clone N418, Thermo Fisher Scientific), CD45.2 (clone 104, BD Bioscience, San Jose, CA), CD45.1 (clone A20, Biolegend) and I-A^b (clone AF6–120.1, Biolegend).

Tanaka S., Gauthier J.M., Terada Y., Takahashi T., Li W., Hashimoto K., Higashikubo R., Hachem R.R., Bharat A., Ritter J.H., Nava R.G., Puri V., Krupnick A.S., Gelman A.E, & Kreisel D. (2020). Bacterial products in donor airways prevent the induction of lung transplant tolerance. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(1), 353-361.

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Multiparametric Flow Cytometry Analysis of Immune Cells

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Cell suspensions were blocked with DPBS/2% FBS containing 5 μg/mL Fc Block (eBioscience) for 1 hr at 4 °C. Blocked cells were stained fluorochrome conjugated monoclonal antibodies in HBSS/3% FBS/0.02% sodium azide, obtained from either BD Bioscience - SIGLECF (E50-2440), CD64 (X54-5/7.1), CD11c (HL3), CD4 (RM4-5), CD8 (53-6.7); or eBioscience - CD45 eFluor 605 (30-F11), MHC class II (I-A/I-E; M5/114.15.2), F4/80 (BM8), Ly-6C (HK1.4); or Biolegend – Ly-6G (1A8), CD11b (M1/70); Biolegend-CD90.2 (clone 30-H12); BB15 CD8, e650 CD4. Cells were washed in DPBS alone and incubated with a Fixable Viability Dye eFluor® 780 (eBioscience) for 20 min at 4 °C to discriminate non-viable cells. Data were collected using a BD LSR flow cytometer and analyzed using FlowJo version 9.8.5 (Treestar, Mountain View, CA) using the gating strategy shown on Supplementary Figure S1. Flow cytometric data were collected using an LSR II (Becton Dickinson) instrument in the Penn State College of Medicine Flow Cytometry Core Facility.

Fino K.K., Yang L., Silveyra P., Hu S., Umstead T.M., DiAngelo S., Halstead E.S., Cooper T.K., Abraham T., Takahashi Y., Zhou Z., Wang H.G, & Chroneos Z.C. (2017). SH3GLB2/endophilin B2 regulates lung homeostasis and recovery from severe influenza A virus infection. Scientific Reports, 7, 7262.

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Single-cell surface marker profiling

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Tissues were processed to single cell suspensions as described [14 (link)]. Cell surface labeling was performed for 20 min at room temperature using antibodies targeting B220 (clone RA3-6B2, Biolegend), CD90.2 (clone 30-H12, Biolegend), CD19 (clone 6D5, Biolegend), CD69 (Clone H1.2F3, Biolegend), CD4 (Clone RM4-5, Biolegend) and CD8 (Clone 53-6.7, Biolegend). Fluorescence Minus One (FMO) controls were performed on pooled cells from all experimental groups to assist gating. When median fluorescence intensities (MFI)s were computed, baseline fluorescence obtained with FMO controls was subtracted. Data were acquired using a FACS Diva-driven customized LSR Fortessa (BD Biosciences, NJ, USA) and analyzed using the FlowJo software (Tree Star, OR, USA).

Courtemanche O., Huppé C.A., Blais Lecours P., Lerdu O., Roy J., Lauzon-Joset J.F., Blanchet M.R., Morissette M.C, & Marsolais D. (2022). Co-modulation of T cells and B cells enhances the inhibition of inflammation in experimental hypersensitivity pneumonitis. Respiratory Research, 23, 275.

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