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Quantum alexafluor 647 mesf calibration beads

Manufactured by Bangs Laboratories
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Quantum AlexaFluor 647 MESF calibration beads are a set of fluorescent particles designed for the calibration of flow cytometers and other fluorescence-based instruments. These beads provide a standardized fluorescence signal intensity, allowing for the quantification of the brightness of fluorescent samples in Molecules of Equivalent Soluble Fluorophore (MESF) units.

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Lab products found in correlation

Anti cd19 pe cy7, by BD (2 mentions) Alexafluor 647 conjugated anti btla, by BioLegend (2 mentions) Anti cd38 alexafluor 488, by BioLegend (2 mentions) Anti hvem alexafluor 647, by BD (2 mentions) Anti cd27 brilliantviolet421, by BioLegend (2 mentions) Anti cd20 pe, by BioLegend (2 mentions)

2 protocols using quantum alexafluor 647 mesf calibration beads

1

Quantification of HVEM Expression on B Cell Subsets

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Isolated whole CD4+ populations or isolated Tfh cells were stained with with AlexaFluor 647-conjugated anti-BTLA (BioLegend, 344520) and compared to Quantum AlexaFluor 647 MESF calibration beads (Bangs Laboratories) by flow cytometry. HVEM levels on B cells were determined by staining PBMCs from whole blood with anti-CD19-PE-Cy7 (BD Biosciences; 560911), anti-CD38-AlexaFluor 488 (BioLegend; 303511), anti-CD27-BrilliantViolet421 (BioLegend; 356417), anti-CD20-PE (BioLegend; 302305), and anti-HVEM-AlexaFluor 647 (BD Biosciences; 564411). B cell subsets were gated as follows: Activated (CD19+ CD20+ CD27+ CD38+), Memory (CD19+ CD20+ CD27+ CD38-), immature/transitional (CD19+ CD20+ CD27- CD38+), and plasmablasts (CD19+ CD20- CD27+ CD38+). HVEM intensity on each cell subset was converted to absolute protein numbers by reference to Quantum AlexaFluor 647 MESF calibration beads (Bangs Laboratories). Based on the mean HVEM expression on activated B cells of ∼35,000/cell, minimum surface density was estimated at ∼35 molecules/μm2 assuming an upper limit of ∼1000 μm2 total area.
Mintz M.A., Felce J.H., Chou M.Y., Mayya V., Xu Y., Shui J.W., An J., Li Z., Marson A., Okada T., Ware C.F., Kronenberg M., Dustin M.L, & Cyster J.G. (2019). The HVEM-BTLA Axis Restrains T Cell Help to Germinal Center B Cells and Functions as a Cell-Extrinsic Suppressor in Lymphomagenesis. Immunity, 51(2), 310-323.e7.
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2

Quantifying HVEM Expression on B Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols
Isolated whole CD4+ populations or isolated Tfh cells were stained with with AlexaFluor 647-conjugated anti-BTLA (BioLegend, 344520) and compared to Quantum AlexaFluor 647 MESF calibration beads (Bangs Laboratories) by flow cytometry. HVEM levels on B cells were determined by staining PBMCs from whole blood with anti-CD19-PE-Cy7 (BD Biosciences; 560911), anti-CD38-AlexaFluor 488 (BioLegend; 303511), anti-CD27-BrilliantViolet421 (BioLegend; 356417), anti-CD20-PE (BioLegend; 302305), and anti-HVEM-AlexaFluor 647 (BD Biosciences; 564411). B cell subsets were gated as follows: Activated (CD19+ CD20+ CD27+ CD38+), Memory (CD19+ CD20+ CD27+ CD38-), immature/transitional (CD19+ CD20+ CD27- CD38+), and plasmablasts (CD19+ CD20- CD27+ CD38+). HVEM intensity on each cell subset was converted to absolute protein numbers by reference to Quantum AlexaFluor 647 MESF calibration beads (Bangs Laboratories). Based on the mean HVEM expression on activated B cells of ~35,000/cell, minimum surface density was estimated at ~35 molecules/µm2 assuming an upper limit of ~1000 µm2 total area.
Mintz M.A., Felce J.H., Chou M.Y., Mayya V., Xu Y., Shui J.W., An J., Li Z., Marson A., Okada T., Ware C.F., Kronenberg M., Dustin M.L, & Cyster J.G. (2019). The HVEM-BTLA Axis Restrains T Cell Help to Germinal Center B Cells and Functions as a Cell-Extrinsic Suppressor in Lymphomagenesis. Immunity, 51(2), 310-323.e7.
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