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Cd140a

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CD140A is a recombinant protein produced in mouse myeloma cell line. It is a member of the platelet-derived growth factor receptor (PDGFR) family and functions as a cell surface receptor for various ligands.

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Lab products found in correlation

Hla dr, by BD (2 mentions) Cd144, by Thermo Fisher Scientific (1 mentions) Cd106, by Thermo Fisher Scientific (1 mentions) Cd146, by Thermo Fisher Scientific (1 mentions) Summit3, by Beckman Coulter (1 mentions) Cd106, by BD (1 mentions) Fc500 analyzer, by Beckman Coulter (1 mentions) Cd140a, by BD (1 mentions) Cd146, by Beckman Coulter (1 mentions) Vegfr1, by R&D Systems (1 mentions) Cd184, by BD (1 mentions) Cd11b, by Beckman Coulter (1 mentions) Fabp4, by Abcam (1 mentions) Rab27a, by R&D Systems (1 mentions) Srx 101a, by Konica Minolta (1 mentions) C ebpα, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions) Pgc 1a, by Abcam (1 mentions) Plin1, by Santa Cruz Biotechnology (1 mentions) Tgx gel, by Bio-Rad (1 mentions)

3 protocols using cd140a

1

Multimarker Immunophenotyping of SVF Cells

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Cells from the SVF at passage 3 were immunophenotyped by flow cytometry using cell surface markers CD14, CD34, CD45, CD73, HLA-DR (BD Pharmigen, EEUU), CD29, CD31, CD44, CD49D, CD19, CD90, CD105, CD106, CD133, CD144, CD146, (eBioscience), CD140A, CD140B, CD166 (RD Systems, EEUU). The clone, fluorochrome and amount of each antibody are provided in S1 Table. Briefly, 106cells/ml were resuspended in blocking buffer solution containing PBS supplemented with 3.0% BSA, and incubated on ice for 10 minutes. Then, they were gated according to their granularity (SS) and size (FS). 1x105 cells were stained using the antibodies (0.25 μg x 106 cells in 100 μl PBS) against the above surface markers or the isotype-matched control antibody. Samples were analyzed on a FACScan CyAnTM High-speed ADP Analyzer (Beckman Coulter, CA. EEUU). Data acquisition and analysis were performed using SUMMIT 3 software (Beckman Coulter). Unstained cells were used to establish flow cytometer settings. Debris and cells/particles with auto-fluorescence were removed by using a threshold on the forward scatter. The original (RAW) set of data is shown in S1 Fig.
Oliva-Olivera W., Coín-Aragüez L., Salas J., Lhamyani S., Gentile A.M., Sarria García E., Hmadcha A., Zayed H., Vega-Rioja A., Tinahones F.J, & El Bekay R. (2015). Myocardial Ischemic Subject’s Thymus Fat: A Novel Source of Multipotent Stromal Cells. PLoS ONE, 10(12), e0144401.
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2

Multiparametric Characterization of MSCs

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A four-color FC analysis was performed on FC500 analyzer (Beckman Coulter) to characterize MSCs and S-MSCs at day 28: FITC-conjugated CD105 (endoglin), CD31 (PECAM1) antibodies and PE-conjugated CD90 (Thy-1), CD140a (Platelet-derived growth factor receptor alpha, PDGF RA), CD140b (Platelet-derived growth factor receptor beta, PDGF RB), CD144 (Ve-Cadherin, CDH5), VEGF R1 (Vascular endothelial growth factor receptor 1, FLT1) antibodies and PE-Cy5-conjugated HLA-DR (human leukocyte antigen-D related), CD34, CD45, CD11b (integrin alpha M), CD146 (Melanoma adhesion molecule, MCAM), CD184 (CXCR4), CD106 (VCAM1) antibodies and PE-Cy7 conjugated CD73 (5′ nucleotidase) and VEGF R2 (KDR, CD309) antibodies. Mouse anti-Human CD105, CD90, CD73, HLA-DR, CD140a, CD31, CD184, CD106 were obtained from BD Biosciences (Le Pont de Claix, France), CD34, CD45, CD11b, CD146, VEGFR2 from Beckman Coulter, CD140a, VEGFR1 from R&D Systems Inc (Minneapolis, USA) and CD144 from Santa-Cruz Biotechnology inc SCBT (Dallas, USA). The isotype-matched mouse IgG1-FITC, IgG1-PE, IgG1-PCy5 and IgG1-PCy7 were used as negative controls. Acquisition and processing data from 7,000 events were analysed using Kaluza software.
Al-Rifai R., Nguyen P., Bouland N., Terryn C., Kanagaratnam L., Poitevin G., François C., Boisson-Vidal C., Sevestre M.A, & Tournois C. (2019). In vivo efficacy of endothelial growth medium stimulated mesenchymal stem cells derived from patients with critical limb ischemia. Journal of Translational Medicine, 17, 261.
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3

Immunoblotting for Adipogenic Markers

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For immunoblotting, 25–50μg of protein was loaded onto a 10% TGX gel (Bio-Rad) and separated at 200 volts for ~30 to 40 minutes. Protein was transferred to a PVDF membrane using Bio-Rad Trans-blot Turbo (1.5amps, 15 volts for 30 minutes). Membranes were blocked using 5% w/v non-milk dissolved in PBS with 0.1% tween-20 (PBS-T) for 30 minutes and incubated with primary antibody overnight at 4°C. Primary antibodies and dilutions used were as follows: from Cell Signaling Technologies - ACTB 1:5000 (#4970S), C/EBPα 1:750 (#8178S), FASN 1:1000 (#3180S), PLIN1 1:4000 (#9349S), PPARG 1:500 (#2443S), TUBA1B 1:2500 (#2125S); from Santa Cruz Biotechnology - CD73 1:1000 (#sc-398260), CD140A 1:1000 (#sc-398206), Rab5 1:750 (#sc130010), Rab27a 1:1000 (#sc136996); from R&D Systems - UCP1 1:750 (#MAB6158-SP); and from Abcam - FABP4 1:2500 (#ab92501), PGC1A 1:2000 (#ab54481). The next day, the membranes were washed 3× for 15 minutes each in PBS-T and incubated in HRP-conjugated secondary antibody from Cell Signaling Technology (#7074S and #7076S) diluted 1:4000 in PBT-T supplemented with 5% w/v milk for 1h. Membranes were washed 3× for 15 minutes each and developed using Immobilon forte chemiluminescent reagent (Millipore cat #WBLUF0100). Signal was detected using HyBlotCL ® autoradiography films (Denville #E3212–1001371) in a dark room and developed on a Konica developer (SRX-101A).
Boucher J.M., Robich M., Scott S.S., Yang X., Ryzhova L., Turner J.E., Pinz I, & Liaw L. (2018). Rab27a regulates human perivascular adipose progenitor cell differentiation. Cardiovascular drugs and therapy, 32(5), 519-530.
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