Anti b220 biotin

Manufactured by BD

Anti-B220-biotin is a laboratory reagent used in flow cytometry applications. It is a biotinylated monoclonal antibody that targets the B220 (CD45R) antigen expressed on the surface of B cells. This product can be used for the identification and isolation of B cells from various sample types.

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Lab products found in correlation

Leica las x software, by Leica camera (2 mentions) Gl7 alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Ki 67 horizon v450, by BD (2 mentions) Tcs spe confocal microscope, by Leica camera (2 mentions) Cryomatrix embedding resin, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594 conjugated streptavidin, by Thermo Fisher Scientific (2 mentions) Cryostar nx70 cryostat, by Thermo Fisher Scientific (2 mentions) Prolong gold antifade mounting media, by Thermo Fisher Scientific (2 mentions) Streptavidin alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Gl7 efluor 660, by Thermo Fisher Scientific (1 mentions) Ki67 v450, by BD (1 mentions) Dako fluorescent mounting medium, by Agilent Technologies (1 mentions) Anti cd43 pe, by BD (1 mentions) Anti cd8a pe, by BD (1 mentions) Anti b220 fitc, by BD (1 mentions) Anti cd21 fitc, by BD (1 mentions) Anti b220 percp cy5, by BD (1 mentions) Anti human igκ fitc, by Southern Biotech (1 mentions) Streptavidin apc, by Southern Biotech (1 mentions) Cellquest software, by BD (1 mentions)

4 protocols using anti b220 biotin

Immunohistochemical Analysis of Germinal Centers

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Three days after the second s.l. immunization, spleens were removed and embedded in Tissue-Tek OCT compound (Sakura, Zoeterwoude, The Netherlands). For some analyses, tissues were immediately frozen in liquid nitrogen and stored at −70°C. 7-μm-thick frozen tissue sections were prepared and then fixed in acetone, air-dried, and rinsed in PBS. Spleen cryosections were stained with anti-B220, GL7, and Ki67 (anti-B220-Biotin, BD #553086 followed by Streptavidin Alexa Fluor 594, Invitrogen #S11227; GL7-eFluor 660, eBioscience #50-5902-82; Ki67-V450, BD #561281). Slides were mounted with Dako fluorescent mounting medium #S3023 (Dako, Glostrup, Denmark). Confocal images were acquired using an inverted LSM 700 Axio Observer.Z1 under Plan-Apochromat 20×/1.4 DIC immersion objective (Carl Zeiss, Jena, Germany).

Sun J.B., Holmgren J., Larena M., Terrinoni M., Fang Y., Bresnick A.R, & Xiang Z. (2017). Deficiency in Calcium-Binding Protein S100A4 Impairs the Adjuvant Action of Cholera Toxin. Frontiers in Immunology, 8, 1119.

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Isolation and Characterization of Murine Immune Cell Subsets

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Single-cell suspensions were prepared from spleen, bone marrow and thymus of 6-14 week old mice. Single-cell suspensions were stained with antibodies and analyzed using FACS Calibur with CellQuest software (BD Bioscence, San Diego, CA) or FlowJo software (Tree Star, Ashland, OR) (15 (link)). B220⁺CD43^-IgM^- small pre-B cells and CD4⁺CD8⁺ double-positive T cells were sorted by a MoFlo flow cytometer. Splenic B cells were purified using B cell isolation kits with MACS cell separation columns (Miltenyi Biotec, San Diego, CA). Generally we pooled bone marrow or splenic cells from 2-3 animals of the same genetic background for cell fractionation. Antibodies used are as follows: anti-mouse-Igκ-PE (BD Bioscience); anti-mouse-Igλ1,2,3-FITC (BD Bioscience); anti-human-Igκ-FITC (Southern Biotech, Birmingham, AL); anti-B220-PerCP-Cy5.5, anti-IgM-APC, anti-CD43-PE, anti-B220-FITC, anti–CD21-FITC, anti–CD23-APC, anti–CD4-FITC, anti–CD8a-PE, anti-B220-biotin (all from BD Bioscience); Streptavidin-APC (Southern Biotech).

Xiang Y., Park S.K, & Garrard W.T. (2014). A major deletion in the Vκ-Jκ intervening region results in hyper-elevated transcription of proximal Vκ genes and a severely restricted repertoire. Journal of immunology (Baltimore, Md. : 1950), 193(7), 3746-3754.

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Multicolor Immunofluorescence Staining of Mouse Spleen

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Mouse spleens were embedded in Cryomatrix^™ embedding resin (Thermo Fisher Scientific). Frozen tissues were sectioned at 7 μm thick using CryoStar^™ NX70 Cryostat (Thermo Fisher Scientific), then fixed in acetone, air-dried, and rinsed in PBS. Next, spleen cryosections were stained with anti-B220-biotin (BD Biosciences), GL7-Alexa Fluor 488 (eBioscience), and Ki-67-Horizon V450 (BD Biosciences), followed by incubation with Alexa Fluor 594-conjugated streptavidin (Invitrogen). Slides were mounted with ProLong^™ gold antifade mounting media (Thermo Fisher Scientific). Confocal images were acquired using a TCS SPE confocal microscope (Leica) with a 60× zoom. The data were analyzed using LAS X Leica software (Leica).

Sen Chaudhuri A., Yeh Y.W., Zewdie O., Li N.S., Sun J.B., Jin T., Wei B., Holmgren J, & Xiang Z. (2022). S100A4 exerts robust mucosal adjuvant activity for co-administered antigens in mice. Mucosal Immunology, 15(5), 1028-1039.

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Immunofluorescent Staining of Mouse Spleen

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Mouse spleens were embedded in Cryomatrix™ embedding resin (Thermo Fisher Scientific). Frozen tissues were sectioned at 7 µm thick using CryoStar™ NX70 Cryostat (Thermo Fisher Scientific), then fixed in acetone, air-dried, and rinsed in PBS. Next, spleen cryosections were stained with anti-B220-biotin (BD Biosciences), GL7-Alexa Fluor 488 (eBioscience), and Ki-67-Horizon V450 (BD Biosciences), followed by incubation with Alexa Fluor 594-conjugated streptavidin (Invitrogen). Slides were mounted with ProLong™ gold antifade mounting media (Thermo Fisher Scientific).
Confocal images were acquired using a TCS SPE confocal microscope (Leica) with a 60 × zoom. The data were analyzed using LAS X Leica software (Leica).

Chaudhuri A.S., Yeh Y., Sun J., Zewdie O., Jin T., Wei B., Holmgren J., & Xiang Z. (2022). S100A4 exerts robust mucosal adjuvant activity for co-administered antigens in mice.

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