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2 protocols using slc27a4

1

Flow Cytometry Immunophenotyping Protocol

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For in-vitro studies the following flow cytometry panel was used in conjunction with CTV and viability dye: anti-CD3 Alexa700, CD4 PE-Cy7, CD8 APC, CD44 PE all at a 1:200 dilution purchased from Biolegend. For in-vivo and ex-vivo studies, the following flow cytometry panel was used: anti-CD3 PE-Cy7, CD4 Pacific Blue, CD8 APC, CD44 PE, and anti-CD25 Alexa 700 all at a 1:200 dilution (except anti-CD25 at a 1:50 dilution) and were purchased from Biolegend. Endogenous Foxp3 expression was detected via eYFP fluorescence (or re-stained with anti-Foxp3 Efluor-450 clone FJK-16 s for any overnight staining (Fisher)), all acquisition was performed using a BD Fortessa flow cytometer. For surface FA transporter measurement, anti-CD36-APC (Biolegend), SLC27A1 (sigma), and SLC27A4 was purchased from Abcam (Cambridge, UK), followed by secondary goat anti-rabbit IgG Alexa-647 (Jackson Immunoresearch, West Grove, PA). Proliferation was analyzed using both FACS DIVA software as well as FlowJo software to determine expansion indexes.
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2

Flow Cytometry Immunophenotyping Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
For in-vitro studies the following flow cytometry panel was used in conjunction with CTV and viability dye: anti-CD3 Alexa700, CD4 PE-Cy7, CD8 APC, CD44 PE all at a 1:200 dilution purchased from Biolegend. For in-vivo and ex-vivo studies, the following flow cytometry panel was used: anti-CD3 PE-Cy7, CD4 Pacific Blue, CD8 APC, CD44 PE, and anti-CD25 Alexa 700 all at a 1:200 dilution (except anti-CD25 at a 1:50 dilution) and were purchased from Biolegend. Endogenous Foxp3 expression was detected via eYFP fluorescence (or re-stained with anti-Foxp3 Efluor-450 clone FJK-16 s for any overnight staining (Fisher)), all acquisition was performed using a BD Fortessa flow cytometer. For surface FA transporter measurement, anti-CD36-APC (Biolegend), SLC27A1 (sigma), and SLC27A4 was purchased from Abcam (Cambridge, UK), followed by secondary goat anti-rabbit IgG Alexa-647 (Jackson Immunoresearch, West Grove, PA). Proliferation was analyzed using both FACS DIVA software as well as FlowJo software to determine expansion indexes.
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