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B220 ra3 6b2 apc

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The B220 (RA3–6B2) APC is a lab equipment product from Thermo Fisher Scientific. It is an antibody-based reagent designed for flow cytometry applications. The core function of this product is to detect and identify B220-positive cells, which are a subset of lymphocytes.

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Lab products found in correlation

Cy3 streptavidin, by Jackson ImmunoResearch (3 mentions) Orca er, by Hamamatsu Photonics (3 mentions) Tissue tek oct, by Sakura Finetek (3 mentions) Cd21 cd35 4e3 fitc, by Thermo Fisher Scientific (2 mentions) Pna biotin, by Vector Laboratories (2 mentions) Myc y69, by Abcam (1 mentions) Igm 2 41 apc, by Thermo Fisher Scientific (1 mentions) Apoptag peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions) Propidium iodide staining, by Merck Group (1 mentions) Cd138 pe 281 2, by BD (1 mentions) Cd138 apc 281 2, by BD (1 mentions) Cyan adp analyzer, by Beckman Coulter (1 mentions)

4 protocols using b220 ra3 6b2 apc

1

Immunophenotyping and Proliferation Analysis

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Cells stimulated with ALD-DNA, LPS, or a combination of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience), CD86-PE (GL1, eBioscience), MHC class II-FITC (2G9, BD Biosciences), or CD138-PE (281-2, BD Biosciences). CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences). Appropriate isotype-matched antibodies were included as controls. Data were acquired on a CyAn ADP Analyzer and analyzed using Summit 5.2 software (both from Beckman-coulter). Dead cells and debris were excluded by forward scatter and side scatter (FSC/SSC) gating and propidium iodide staining (Sigma-Aldrich). For BrdU incorporation assay using a FITC BrdU Flow kit, cells were fixed, permeabilized, and treated with DNase prior to BrdU staining using a FITC-coupled anti-BrdU monoclonal antibody, as recommended by the manufacturer. The incorporated BrdU was measured in conjunction with DNA content by 7-AAD staining.
Lu Q., Wang J.Y., Wang L., Jiang X, & Chu Y. (2014). Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production. PLoS ONE, 9(10), e109095.
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2

Immunofluorescence Analysis of Splenic Tissue

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Spleens embedded in Tissue TEK O.C.T. (Sakura Finetek) were sectioned on a Microtome Cryostat HM 505 E (Microm). Sections were fixed with acetone and blocked with 5% FBS in PBS. Fluorescent images were acquired on an Axio Imager.M1 (Zeiss) microscope equipped with an Orca-ER (Hamamatsu) camera. SlideBook (3i) was used as the imaging software. Gimp (GNU Image Manipulation Program) was used for image editing. The following antibodies/reagents were used: B220 (RA3–6B2) APC, CD21/CD35 (4E3) FITC (Thermo Fisher Scientific), PNA Biotin (Vector Laboratories) and Streptavidin-Cy3 (Jackson ImmunoResearch Labs).
Ramezani-Rad P., Chen C., Zhu Z, & Rickert R.C. (2020). Cyclin D3 Governs Clonal Expansion of Dark Zone Germinal Center B Cells. Cell reports, 33(7), 108403.
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3

Multicolor Immunofluorescence Analysis of Mouse Spleen

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Spleens embedded in Tissue TEK O.C.T. (Sakura Finetek) were sectioned on a Microtome Cryostat HM 505 E (Microm). Sections were fixed with acetone and blocked with 5% FBS in PBS. Florescent images were acquired on an Axio Imager.M1 (Zeiss) microscope equipped with an Orca-ER (Hamamatsu) camera. SlideBook (3i) was used as the imaging software. Gimp (GNU Image Manipulation Program) was used for image editing. The following antibodies were used: B220 (RA3-6B2) APC, IgM (II/41) APC, IgD (11-26c) PE (Thermo Fisher Scientific), Moma1 Biotin (Abcam), and Streptavidin-Cy3 (Jackson ImmunoResearch). For immunohistochemical analysis, spleens were formalin-fixed and paraffin-embedded. B220 (RA3-6B2) Biotin (BD Biosciences) and Myc (Y69) (Abcam) and were used as primary antibodies and horseradish-peroxidase-coupled Streptavidin or anti-Rabbit IgG were used as secondary antibodies. TUNEL staining was performed by using the ApopTag Peroxidase In Situ Apoptosis Detection Kit (MilliporeSigma) according to the manufacturer’s protocol.
Ramezani-Rad P., Leung C.R., Apgar J.R, & Rickert R.C. (2020). E3 ubiquitin ligase Fbw7 regulates the survival of mature B cells. Journal of immunology (Baltimore, Md. : 1950), 204(6), 1535-1542.
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4

Immunofluorescence Analysis of Splenic Tissue

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Spleens embedded in Tissue TEK O.C.T. (Sakura Finetek) were sectioned on a Microtome Cryostat HM 505 E (Microm). Sections were fixed with acetone and blocked with 5% FBS in PBS. Fluorescent images were acquired on an Axio Imager.M1 (Zeiss) microscope equipped with an Orca-ER (Hamamatsu) camera. SlideBook (3i) was used as the imaging software. Gimp (GNU Image Manipulation Program) was used for image editing. The following antibodies/reagents were used: B220 (RA3–6B2) APC, CD21/CD35 (4E3) FITC (Thermo Fisher Scientific), PNA Biotin (Vector Laboratories) and Streptavidin-Cy3 (Jackson ImmunoResearch Labs).
Ramezani-Rad P., Chen C., Zhu Z, & Rickert R.C. (2020). Cyclin D3 Governs Clonal Expansion of Dark Zone Germinal Center B Cells. Cell reports, 33(7), 108403.
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