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Alexafluor 647 conjugated anti btla

Manufactured by BioLegend

AlexaFluor 647-conjugated anti-BTLA is a fluorescently labeled antibody that binds to the BTLA (B and T Lymphocyte Attenuator) protein. It can be used for the detection and analysis of BTLA expression in various cell types.

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2 protocols using alexafluor 647 conjugated anti btla

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.
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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.
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