Anti mouse igg2b antibody staining

Manufactured by Thermo Fisher Scientific

The Anti–mouse IgG2b antibody is a laboratory reagent used to detect and measure the presence of mouse immunoglobulin G subclass 2b (IgG2b) in biological samples. It is a key tool for immunoassays and other immunological techniques.

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Lab products found in correlation

Alexa fluor 450 streptavidin, by Thermo Fisher Scientific (2 mentions) Soluble biotinylated anti igm, by Southern Biotech (2 mentions) Ebioy ae, by Thermo Fisher Scientific (2 mentions)

2 protocols using anti mouse igg2b antibody staining

B Cell Antigen Internalization and Presentation

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For internalization assays, purified B cells were loaded with IgM coated beads or with soluble biotinylated anti-IgM (Southern Biotech) on ice for 30 min. Cells were then washed with PBS 2% FCS to remove excess antigen, and then incubated for 5–30 min at 37°C. After fixation with 2% paraformaldehyde, noninternalized beads were detected with Alexa Fluor 450 streptavidin (eBioscience).
To detect antigen presentation, B cells loaded with Ea peptide and IgM-coated beads were incubated between 3 and 5 h at 37°C, and then fixed in 2% formaldehyde. These cells were then stained with anti-MHCII/Ea antibody (eBioY-Ae; eBioscience), followed by anti–mouse IgG2b antibody staining (Life Technologies) for detection.

Burbage M., Keppler S.J., Gasparrini F., Martínez-Martín N., Gaya M., Feest C., Domart M.C., Brakebusch C., Collinson L., Bruckbauer A, & Batista F.D. (2015). Cdc42 is a key regulator of B cell differentiation and is required for antiviral humoral immunity. The Journal of Experimental Medicine, 212(1), 53-72.

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Internalization and Antigen Presentation Assay for B Cells

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For internalization assays, purified B cells were loaded with IgM coated fluorescently-labelled microspheres or with soluble biotinylated anti-IgM (Southern Biotech) on ice for 30 min. Cells were then washed with PBS 2% FCS to remove excess antigen and then incubated for 5 to 30 min at 37°C. After fixation with 2% paraformaldehyde, non-internalised beads or anti-IgM were detected with AlexaFluor450 streptavidin (eBioscience, San Diego, California). In case of bead internalisation, bead-positive cells were selected on the basis of the bead fluorescence.
To detect antigen presentation, B cells loaded with Eα peptide and IgM coated microspheres were incubated between 3 and 5 hr at 37°C and then fixed in 2% formaldehyde. These cells were then stained with anti-MHCII/Eα antibody (eBioY-Ae, eBioscience), followed by anti-mouse IgG2b antibody staining (Life Technologies) for detection.

Burbage M., Gasparrini F., Aggarwal S., Gaya M., Arnold J., Nair U., Way M., Bruckbauer A, & Batista F.D. (2018). Tuning of in vivo cognate B-T cell interactions by Intersectin 2 is required for effective anti-viral B cell immunity. eLife, 7, e26556.

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