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Anti b220 cd45r

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Anti-B220 (CD45R) is a mouse monoclonal antibody that recognizes the B220 (CD45R) cell surface antigen. B220 (CD45R) is a protein tyrosine phosphatase that is expressed on the surface of B cells and a subset of T cells. The antibody can be used for the identification and isolation of B cells in flow cytometry and other immunological applications.

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Lab products found in correlation

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Close spelling variants of this product

Anti-B220 (40 citations) CD45R/B220 (19 citations) Anti-CD45R (4 citations) Anti-CD45R/B220 (clone RA3-6B2, (4 citations) APC anti-mouse/human CD45R/B220 (3 citations) Alexa-488 anti-CD45R (B220) (clone RA3–6B2), (2 citations) Anti-CD45R(B220)-eFluor780 (2 citations) Biotin anti-CD45R (B220) (RA3-6B2) (2 citations) FITC-rat anti-mouse/human CD45R/B220 (clone RA3–6B2, (2 citations) Alexa Flour 700 anti-mouse CD45R/B220 (RA3-6B2, (2 citations)

5 protocols using anti b220 cd45r

1

Multiparametric Flow Cytometry for Immune Cell Analysis

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To determine chimerism, splenocytes were stained with anti‐H‐2Dd‐FITC (clone: 34–2–12) and anti‐H‐2Db‐PE (clone: KH95) (all from Biolegend). Treg cells were determined in the spleens according to manufacturer's instructions using a kit from eBioscience (San Diego, CA). Spleen cells were also characterized using anti‐CD11b (clone: M1/70), anti‐CD11c (clone: N418), anti‐B220/CD45R (clone: RA3‐6B2), anti‐I‐A/I‐E (clone: M5/114.152), anti‐CD4 (clone: RM4‐5 and GK1.5), anti‐pDCA1 (clone: 927), anti‐CD8α (clone: 100712), anti‐TCR‐β (clone: H57‐597), all from Biolegend, and a viability marker (LIVE/DEAD® Fixable Near‐IR Dead Cell Stain Kit, Life Technologies, Eugene, OR). For MACS cell purification, we used anti‐CD11c (cat. 130‐052‐001), anti‐CD4 (cat. 130‐049‐201), and anti‐CD8 (cat. 130‐049‐301) microbeads (Miltenyi Biotec Bergisch Glabach, Germany). For the in vitro functional assays, DC subpopulations were purified from spleens by MACS. CD11c cells were then sorted by FACS (BD FacsAria III) using APC/Cy7‐labeled anti‐B220, PercP‐labeled anti‐I‐A/I‐E, APC‐labeled anti‐pDCA1, PE‐labeled anti‐CD11b and PE/Cy7‐CD8a. We obtained 90.9 ± 1.35% purity for CD8αcDCs, 95.1 ± 1% for CD11b cDCs and 94.± 0.3% for pDCs. All cells were acquired using a FACS‐LSRFortessa according to BD bioscience protocols and analyzed by FlowJo software version 9.8.1.
Gaignage M., Marillier R.G., Uyttenhove C., Dauguet N., Saxena A., Ryffel B., Michiels T., Coutelier J, & Van Snick J. (2017). Mouse nidovirus LDV infection alleviates graft versus host disease and induces type I IFN‐dependent inhibition of dendritic cells and allo‐responsive T cells. Immunity, Inflammation and Disease, 5(2), 200-213.
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2

Multiparametric Flow Cytometry of Lymphocytes

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Cells suspended in PBS supplemented with 0.5% w/v bovine albumin serum (BSA) and 2 mM EDTA were stained with combinations of the following antibodies: anti-CD19 (BD Biosciences, BV421), anti-GL7 (eBioscience, Alexa488), anti-Fas (CD95) (eBioscience, APC), anti-B220 (CD45R) (BioLegend, BV711 & APC, BD Biosciences, BUV496), anti-CD38 (BD Biosciences, BUV395), anti-CXCR4 (eBioscience, PE/Cy7), anti-CD86 (BD Biosciences, BV421, eBiosciences, FITC), anti-IgG1 (eBioscience, PE/Cy7), anti-CD69 (BioLegend, APC), anti-IgM (BD Biosciences, BV711), and anti-MHCII (eBioscience, PE/Cy5). Typically, 0.06–1 μg of antibody was used for staining of approximately 105–107 cells in 100 μL. All samples were analyzed using a BD Accuri C6 or BD LSRFortessa (BD Biosciences, Franklin Lakes, NJ, USA). The data were analyzed using FlowJo software.
Kramer L., Song H.W., Mitchell K., Kartik M., Jain R., Escarra V.L., Quiros E., Fu H., Singh A, & Roy K. (2022). Lipid Membrane-Based Antigen Presentation to B Cells Using a Fully Synthetic Ex Vivo Germinal Center Model. Advanced nanobiomed research, 2(7), 2100137.
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3

Multiparametric Flow Cytometry of Lymphocytes

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Cells suspended in PBS supplemented with 0.5% w/v bovine albumin serum (BSA) and 2 mM EDTA were stained with combinations of the following antibodies: anti-CD19 (BD Biosciences, BV421), anti-GL7 (eBioscience, Alexa488), anti-Fas (CD95) (eBioscience, APC), anti-B220 (CD45R) (BioLegend, BV711 & APC, BD Biosciences, BUV496), anti-CD38 (BD Biosciences, BUV395), anti-CXCR4 (eBioscience, PE/Cy7), anti-CD86 (BD Biosciences, BV421, eBiosciences, FITC), anti-IgG1 (eBioscience, PE/Cy7), anti-CD69 (BioLegend, APC), anti-IgM (BD Biosciences, BV711), and anti-MHCII (eBioscience, PE/Cy5). Typically, 0.06–1 μg of antibody was used for staining of approximately 105–107 cells in 100 μL. All samples were analyzed using a BD Accuri C6 or BD LSRFortessa (BD Biosciences, Franklin Lakes, NJ, USA). The data were analyzed using FlowJo software.
Kramer L., Song H.W., Mitchell K., Kartik M., Jain R., Escarra V.L., Quiros E., Fu H., Singh A, & Roy K. (2022). Lipid Membrane-Based Antigen Presentation to B Cells Using a Fully Synthetic Ex Vivo Germinal Center Model. Advanced nanobiomed research, 2(7), 2100137.
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4

Murine Hematopoietic Stem Cell Isolation

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Female C57BL/6 (B6) and BALB/c mice at 8–10 weeks of age were purchased from SLAC Ltd. Animals were maintained in the Guangxi Normal University Laboratory Animal Center and handled in accordance with the institution’s guidelines. All experimental protocols were approved by the Guangxi Animal Management Committee of Guangxi S&T Department (Project Number syxk (Gui) 2013-0001). Fluorescence-conjugated anti-mouse mAbs anti-c-kit(ACK2), anti-Sca-1(D7), anti-CD34(RAM34), anti-IL-7R(A7R34), anti-FcR II/III(93), anti-Flt3(A2F10), anti-Thy-1.1(HIS51), and their isotype ctrl antibodies (except those for anti-IL-7R and anti-Thy-1.1) were obtained from affymetrix eBioscience. Anti-mouse Lineage cocktail (containing anti-CD3, Ly-6G/Ly-6C, anti-CD11b, anti-CD45R/B220 and anti-TER-119) with isotype ctrl was from Biolegend. Mitomycin C was obtained from Solarbio.
Pu S., Qin B., He H., Zhan J., Wu Q., Zhang X., Yang L., Qu C, & Zhou Z. (2016). Identification of early myeloid progenitors as immunosuppressive cells. Scientific Reports, 6, 23115.
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5

Comprehensive Multiparameter Flow Cytometry

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Flow cytometry was performed using the BD FACSCanto II and analysed with FlowJo software (Tree Star, Ashland, OR). Dead cells were excluded through Fixable Viability Dye eFluor staining (eBioscience). Nonspecific antibody binding was blocked with an anti-CD16/32 (Mouse Fcγ block clone 2.4G2). The cells were incubated for 30 min in 100 μl PBS with conjugated antibodies (BD Biosciences, France): anti CD45 (V500 conjugated; clone 30-F11), CD11b (BV421 conjugated; clone M1/70), anti CD3 (FITC conjugated; clone 145-2C11), anti CD45R/B220 (PE-Cy7 conjugated; clone RA3-6B2), anti Ly6C (FITC conjugated; clone AL-21), anti CD31 (PE conjugated; clone MEC13.3), anti CD34 (Alexa Fluor 647 conjugated; clone RAM34), anti CD146 (FITC conjugated; clone ME-9F1), anti CD29 (PE conjugated; clone HM B1.1), anti Sca-1 (PE-Cy7 conjugated; clone D7), anti CD44 (PE-Cy5.5 conjugated; clone IM7), anti CD105 (Alexa Fluor 647 conjugated; clone MJ7/18), anti SSEA (BV421 conjugated; clone MC631), anti CX3CR1 from Biolegend (France) (PE-Cy7 conjugated; clone SA011F11), anti CCR2 (Alexa Fluor 647 conjugated; clone SA203G11) and CD90 from southern Biotech (FITC conjugated; clone SA203G11). Isotype-matched antibodies (BD Biosciences) were used for control staining. All antibodies were used at 1:100 dilution. The concentration of cell suspensions was adjusted to 1 × 106 cells per 100 μl.
Bensemmane L., Squiban C., Demarquay C., Mathieu N., Benderitter M., Le Guen B., Milliat F, & Linard C. (2021). The stromal vascular fraction mitigates radiation-induced gastrointestinal syndrome in mice. Stem Cell Research & Therapy, 12, 309.
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