For FACS of B cells, PBMC were stained with BUV395 anti-CD19 (BD Biosciences), APC anti-CD27 (Biolegend) and either PE anti-CXCR3 (eBioscience), PE-Cy7 anti-CXCR5 (eBioscience), PE anti-CD11a (Biolegend), PE anti-CD49d (Biolegend), PE anti-CD166 (BD) or FITC anti-CD6 (Biolegend). BUV395 anti-CD19 APC anti-CD27-stained cells were also fixed and permeabilized and stained with rabbit anti-AHNAK (Sigma) and PE donkey anti-rabbit IgG. FCS files of stained cells were collected on a LSRII cytometer (BD) and analyzed using FloJo.
Buv395 anti cd19
Manufactured by BD
The BUV395 anti-CD19 is a fluorescent-labeled monoclonal antibody that binds to the human CD19 antigen. It is designed for use in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd166 pe, by BD (2 mentions)
Anti cd27 apc, by BioLegend (2 mentions)
Anti cd49d pe, by BioLegend (2 mentions)
Lsrii cytometer, by BD (2 mentions)
Pe anti cd11a, by BioLegend (2 mentions)
Pe anti cxcr3, by Thermo Fisher Scientific (2 mentions)
Pe cy7 anti cxcr5, by Thermo Fisher Scientific (2 mentions)
Fitc anti cd6, by BioLegend (2 mentions)
Buv395 anti cd19 apc anti cd27, by Merck Group (2 mentions)
Rabbit anti ahnak, by Merck Group (2 mentions)
Anti cd38 apc, by BD (1 mentions)
Facsaria fusion, by BD (1 mentions)
Telomere pna kit fitc for flow cytometry, by Agilent Technologies (1 mentions)
Rosettesep human b cell enrichment cocktail, by STEMCELL (1 mentions)
Anti cd27 bv785, by BioLegend (1 mentions)
Anti cd11c bv421, by BD (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Lsrii instrument, by BD (1 mentions)
Live dead fixable near ir, by Thermo Fisher Scientific (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
4 protocols using buv395 anti cd19
1
FACS Analysis of B Cell Subsets
2
FACS Analysis of B Cell Subsets
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For FACS of B cells, PBMC were stained with BUV395 anti-CD19 (BD Biosciences), APC anti-CD27 (Biolegend) and either PE anti-CXCR3 (eBioscience), PE-Cy7 anti-CXCR5 (eBioscience), PE anti-CD11a (Biolegend), PE anti-CD49d (Biolegend), PE anti-CD166 (BD) or FITC anti-CD6 (Biolegend). BUV395 anti-CD19 APC anti-CD27-stained cells were also fixed and permeabilized and stained with rabbit anti-AHNAK (Sigma) and PE donkey anti-rabbit IgG. FCS files of stained cells were collected on a LSRII cytometer (BD) and analyzed using FloJo.
3
Sorting and Analyzing B Cell Subsets
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B cells were enriched from PBMC from either individual healthy donors or two pooled healthy donors (to increase cell yield) with RosetteSep human B cell enrichment cocktail (StemCell Technologies#15064). The enriched B cells were stained as described above with 5 µl anti-CD19 BUV395, 1 µl anti-CD38 APC, 5 µl anti-CD11c BV421 (All from BD Biosciences) and 10 µl anti-CD27 BV785 (BioLegend) per 1 × 106 cells. Using a FACS Aria Fusion (BD Biosciences), CD19+CD11c− B cells were sorted as naive (CD27−CD38+), memory (CD27+CD38−/CD38+), or plasma cells (CD27hiCD38hi). CD19+CD11c+ B cells were sorted as CD27−CD38− B cells. Post sort purity was greater than 90%; B cells from five independent donors were examined. In one of the sorts insufficient cells were obtained from CD11c+ B cells and plasma cells to extract data. The RTL of sorted B cell populations were determined using Telomere PNA Kit/FITC for flow cytometry (Dako #K5327), per manufacturer’s instructions.
4
NK Cell Activation Assay Protocol
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Cells were stained for surface markers for 20 min at room temperature in PBS (Sigma). Extracellular antibodies used were as follows: LIVE/DEAD Fixable Near-IR (Thermo Fischer Scientific; L10119), Anti-CD3-BUV737 (BD Biosciences; 612750), Anti-CD14-BUV737 (BD Biosciences; 612763); Anti-CD14-BUV395 (BD Biosciences; 563561); Anti-CD19-BUV737 (BD Biosciences; 564303), Anti-CD19-BUV395 (BD Biosciences; 563549); Anti-CD56-PerCP-Cy5.5 (Biolegend; 304626), Anti-CD16-Pe-Cy7 (Biolegend; 302016), Anti-CD107a-BV510 (Biolegend; 328632), Anti-CD69-BV650 (Biolegend; 310934). Samples were then washed and fixed using 4% formaldehyde or BD Cytofix/Cytoperm Kit (BD biosciences) according to manufacturer’s directions. Flow cytometry analysis was performed on a LSRII instrument (BD Biosciences). A total of 50,000 to 250,000 events were acquired and analyzed using FlowJo software. The analysis was performed on gated cells that fell within the lymphocyte population, stained as live cells using Live/Dead stain. Cells were then gated for negative CD3/CD14/CD19 expression to exclude other cell populations, and NK cells were identified using CD56/CD16 gating strategy. Gating strategies are shown in Supplementary Figure S1
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