Pe cy7 conjugated anti b220

Manufactured by Thermo Fisher Scientific

Sourced in United States

PE-Cy7-conjugated anti-B220 is a fluorescently labeled antibody that binds to the B220 surface antigen expressed on B cells. It can be used in flow cytometry applications for the identification and analysis of B cell populations.

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3 protocols using pe cy7 conjugated anti b220

Multiparametric Flow Cytometry of Mouse B Cell Subsets

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Flow cytometry analysis of mouse NB and GCB was performed using the following fluorescent-labeled anti–mouse antibodies: APC-conjugated anti-B220 (BD Pharmingen), PE-Cy7-conjugated anti-CD95 (BD Pharmingen), FITC-conjugated anti-GL7 (BD Pharmingen) and PE-conjugated CXCR4 (eBioscience). Cell cycle analysis was performed using the BrdU Flow Kit (BD Pharmingen) and antigen-specific GCB (NP⁺GL7⁺CD95⁺B220⁺) were detected using PE-conjugated NP (biosearch Technologies, Inc.). Ex vivo stimulated B cells were stained with PE-Cy7-conjugated anti-B220 (eBioscience), PE-conjugated anti-IgD (BD Pharmingen) and APC-conjugated anti-IgG1 (BD Pharmingen). DAPI was used for the exclusion of dead cells. Data was acquired on a MACSQuant Analyzer (Miltenyi Biotec) and analyzed using FlowJo 7.6.4 software (TreeStar).

Dominguez P.M., Teater M., Chambwe N., Kormaksson M., Redmond D., Ishii J., Vuong B., Chaudhuri J., Melnick A., Vasanthakumar A., Godley L.A., Papavasiliou F.N., Elemento O, & Shaknovich R. (2015). DNA Methylation Dynamics of Germinal Center B Cells Are Mediated by AID. Cell reports, 12(12), 2086-2098.

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Hematopoietic Stem Cell Transplantation

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C57BL/6 J (CD45.2) and B6.SJL-PtprcaPepcb/BoyJ mice (B6.SJL, CD45.1) were provided by the Animal Centre of the Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC. Sex- and age-matched mice were used and maintained in the SPF-certified animal facility in the same center. The procedures for the animal experiments were approved by the Animal Care and Use Committee at the institutions involved in this study. After transplantation, the mice were raised in sterile squirrel cages in a positive pressure room. All of the antibodies used in this paper were obtained from eBioscience (San Diego, CA, USA), including PE-, FITC-, or PerCP-Cy5.5-conjugated anti-CD45.1, PE-conjugated anti-CD45.2, PE-conjugated anti-Sca-1, APC-conjugated anti-c-Kit, PE-conjugated anti-CD3, PE-Cy7-conjugated anti-B220, APC-conjugated anti-Mac-1, biotin-conjugated lineage markers (anti-CD3, −CD4, −CD8, −B220, −CD11b, −Gr-1, and -Ter119) and streptavidin-PE-Cy7. Neutralizing antibodies against MIP-3β and CCR7 were purchased from R&D Systems (Minneapolis, MN, USA).

Ma S., Shi Y., Pang Y., Dong F., Cheng H., Hao S., Xu J., Zhu X., Yuan W., Cheng T, & Zheng G. (2014). Notch1-induced T cell leukemia can be potentiated by microenvironmental cues in the spleen. Journal of Hematology & Oncology, 7, 71.

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Multiparametric Flow Cytometry Analysis

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Single-cell suspensions from BM and spleen were treated with Gey’s solution to remove red blood cells and resuspended in PBS supplemented with 2% BSA. The cells were then stained with a combination of fluorescence-conjugated antibodies. APC-conjugated anti-B220, anti-IgM, anti-CD4 and anti-CD44, and PE-Cy7-conjugated anti-B220, anti-IFNγ, anti-CD25 and anti-CD23 were purchased from eBioscience. PE-conjugated anti-CD43, anti-CD21, anti-Thy1.2, anti-CD5, anti-CD8, and anti-IgD were purchased from BD Biosciences. Samples were applied to a flow cytometer (LSRII, Becton Dickinson). Data were collected and analyzed using FACSDiva software (Becton Dickinson) or FlowJo software (Tree Star).

Chen Y., Zheng Y., You X., Yu M., Fu G., Su X., Zhou F., Zhu W., Wu Z., Zhang J., Wen R, & Wang D. (2016). Kras is critical for B cell lymphopoiesis. Journal of immunology (Baltimore, Md. : 1950), 196(4), 1678-1685.

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