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Clone wm59

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The Clone WM59 is a monoclonal antibody specific for the human melanoma-associated antigen MCAM/CD146. It can be used for the detection and quantification of MCAM/CD146-expressing cells by flow cytometry.

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Lab products found in correlation

Facsdiva v6, by BD (2 mentions) Zombie violet fixable viability kit, by BioLegend (2 mentions) Clone p1h12, by BioLegend (2 mentions) Facscanto 2, by BD (2 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Clone 12g5, by BioLegend (1 mentions) Clone 2d1, by BioLegend (1 mentions)

3 protocols using clone wm59

1

Isolation and Quantification of FAP and SC

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Muscle specimens underwent digestion separately at 37°C in 0.2% collagenase for 90 minutes followed by 0.4% dispase for 30 minutes. The resulting suspensions were filtered through a 40 μm cell strainer, then pelleted and resuspended in FACS buffer (5% Fetal Bovine Serum in PBS). FAP and SC quantity were reported as a percentage of nondebris live cells, as determined using LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (ThermoFisher) and their respective surface markers. Cells were sorting and quantified using FACS BD Aria II. FAPs were defined as CD31‐/CD45‐/CD29‐/CD56‐/PDGFRα+/CD184‐, and SCs were defined as CD31‐/CD45‐/CD29+/CD56+/PDGFRα‐/CD184+ as previously described.29, 30 Gates were set based on each antibodies' fluorescence minus one for every run. The antibodies used for FACS are as follows at concentration of 1 μg/mL per 1 × 106 cells/mL: CD56 (Biolegend, Clone HCD56, #318304, FITC), CD29 (BD bioscience, Clone MAR4, #743783, Pacific Blue), CD184 (Biolegend, Clone 12G5, #306510, APC), PDGFRα (Biolegend, Clone 16A1, #323508, APC/CY7), CD31 (Biolegend, Clone WM59, #303122, BV605), and CD45 (Biolegend, Clone 2D1, #368524, BV605).
Agha O., Mueller‐Immergluck A., Liu M., Zhang H., Theologis A.A., Clark A., Kim H.T., Liu X., Feeley B.T, & Bailey J.F. (2020). Intervertebral disc herniation effects on multifidus muscle composition and resident stem cell populations. JOR Spine, 3(2), e1091.
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2

Fetal Pericyte Surface Characterization

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Surface staining of fetal pericytes was performed with CD31-AF488 (1:100 dilution; clone WM59, BioLegend, #303110) and CD146-AF647 (1:100 dilution; clone P1H12, BioLegend, #361014) antibodies. Zombie Violet fixable viability kit (BioLegend, #423113) was used for fixable live/dead staining as per manufacturer instructions. FACS acquisitions was performed on BD FACSCanto II (BD Biosciences), using BD FACSDIVA v 6.0. All FACS data were analyzed using FlowJo v10.0 software.
Chen W., Foo S.S., Hong E., Wu C., Lee W.S., Lee S.A., Evseenko D., Lopes Moreira M.E., García-Sastre A., Cheng G., Nielsen-Saines K., Brasil P., Avvad-Portari E, & Jung J.U. (2021). Zika virus NS3 protease induces bone morphogenetic protein-dependent brain calcification in human fetuses. Nature microbiology, 6(4), 455-466.
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3

Fetal Pericyte Surface Characterization

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Surface staining of fetal pericytes was performed with CD31-AF488 (1:100 dilution; clone WM59, BioLegend, #303110) and CD146-AF647 (1:100 dilution; clone P1H12, BioLegend, #361014) antibodies. Zombie Violet fixable viability kit (BioLegend, #423113) was used for fixable live/dead staining as per manufacturer instructions. FACS acquisitions was performed on BD FACSCanto II (BD Biosciences), using BD FACSDIVA v 6.0. All FACS data were analyzed using FlowJo v10.0 software.
Chen W., Foo S.S., Hong E., Wu C., Lee W.S., Lee S.A., Evseenko D., Lopes Moreira M.E., García-Sastre A., Cheng G., Nielsen-Saines K., Brasil P., Avvad-Portari E, & Jung J.U. (2021). Zika virus NS3 protease induces bone morphogenetic protein-dependent brain calcification in human fetuses. Nature microbiology, 6(4), 455-466.
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