Frozen sections of spleens (4 μm) were incubated with PE‐conjugated anti‐CD4 (RM4‐5, BioLegend), Alexa Fluor 488‐conjugated anti‐GL7 (GL7, BD) and Alexa Fluor 647‐conjugated anti‐B220 (RA3‐6B2, BD). Frozen sections of kidneys (4 μm) were stained with goat anti‐mouse IgG (Sigma‐Aldrich, Darmstadt, Germany), FITC rat anti‐mouse C3 (Santa Cruz Biotechnology, Dallas, TX, USA), donkey anti‐goat CY3 (Proteintech, Wuhan, China), Alexa Fluor 647‐conjugated IgG1, PE‐conjugated IgG2a (BioLegend), biotin anti‐IgG2b (BioLegend), biotin anti‐IgG3 (BioLegend) and PE streptavidin. Nuclei were identified by DAPI staining. Images were obtained with a laser scanning confocal microscope (Zeiss, Oberkochen, Germany) and analysed by Photoshop CS4 software (Adobe, San Jose, CA, USA).
Biotin anti igg2b
Manufactured by BioLegend
Sourced in China
Biotin anti‐IgG2b is a lab equipment product that functions as a detection reagent. It is used to bind and label IgG2b antibodies, allowing their identification and quantification in various immunoassays and research applications.
Automatically generated - may contain errors
2 protocols using biotin anti igg2b
1
Immunostaining of Spleens and Kidneys
2
Primary B Cell Phenotyping and Proliferation
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Primary B cells were washed with PBS and stained with B220-FITC and biotin anti-IgG1 (BD), biotin anti-IgG2b (BioLegend), and PE anti-IgA (SouthernBiotech). For biotinylated primary antibodies, cells were then stained with PE-Streptavidin (Beckman Coulter). Data were collected on a BD FACSCanto and analyzed using FlowJo software. At least 20,000 events of live lymphoid cells were recorded. For CFSE staining, freshly isolated primary B cells were washed and resuspended in 0.1% BSA/PBS at 1 × 107 cells/ml and labeled with CFSE at a final concentration of 5 uM for 10 min at 37°C. CFSE was quenched with ice-cold RPMI 1640 medium containing 10% FCS and washed twice with BCM. Labeled cells were then cultured in BCM and appropriate stimuli for 72 or 96 hr days prior to analysis.
For cell cycle analysis, B cells were harvested 72 hr post-stimulation, washed once with PBS, and resuspended in ice-cold 70% ethanol while slowly vertexing. Cells were fixed overnight, then washed with PBS one time. Cell pellets were resuspended in propidium iodide (PI) staining solution (50 μg/ml PI and 100 units/ml RNase A in PBS), then incubated at RT in the dark for 2 hr. 50,000 gated events were collected on a BD FACSCanto and analyzed using FlowJo software.
For cell cycle analysis, B cells were harvested 72 hr post-stimulation, washed once with PBS, and resuspended in ice-cold 70% ethanol while slowly vertexing. Cells were fixed overnight, then washed with PBS one time. Cell pellets were resuspended in propidium iodide (PI) staining solution (50 μg/ml PI and 100 units/ml RNase A in PBS), then incubated at RT in the dark for 2 hr. 50,000 gated events were collected on a BD FACSCanto and analyzed using FlowJo software.
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