Pdgfrα

Manufactured by Thermo Fisher Scientific

Sourced in United States

PDGFRα is a cell surface receptor tyrosine kinase that binds to Platelet-Derived Growth Factor (PDGF). It plays a role in cellular proliferation, survival, and differentiation. The PDGFRα gene encodes the alpha subunit of the PDGF receptor.

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Lab products found in correlation

Pdgfrβ, by Thermo Fisher Scientific (2 mentions) Facsverse, by BD (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (2 mentions) Glial fibrillary acidic protein (gfap), by Agilent Technologies (2 mentions) Fluorsave, by Merck Group (2 mentions) Olig2, by Merck Group (2 mentions) Edu detection kit, by Thermo Fisher Scientific (2 mentions) Nanog, by R&D Systems (2 mentions) Oct3 4, by Santa Cruz Biotechnology (2 mentions) Pdgfrα, by Cell Signaling Technology (2 mentions) Caspr, by Abcam (2 mentions) Human gfap, by BioLegend (2 mentions) Neurofilament h, by BioLegend (2 mentions) Ly6g fitc, by Thermo Fisher Scientific (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Cd11b pe cy7, by Thermo Fisher Scientific (1 mentions) Gr 1 pecy7, by Thermo Fisher Scientific (1 mentions) α sma, by Merck Group (1 mentions) Cd25 apc, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Anti‐PDGFRα (4 citations) PDGFRα-APC (3 citations) Anti-PDGFRα-APC (2 citations) PDGFRα (CD140a)-APC (2 citations) Primary antibodies against PDGFRα (2 citations) Anti-PDGFRα (CD140a)-APC (2 citations) PDGFRα (CD140a), (2 citations)

13 protocols using pdgfrα

UAMC-1110 Immunotherapy Protocol

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UAMC-1110 was generously provided by Dr. Pieter Van der Veken (University of Antwerp). Unless otherwise specified, UAMC-1110 was administered in high molecular weight PEG at a concentration of 20mg/kg by oral gavage twice per day. Fluorescently-conjugated antibodies CD3-e450, CD8-PerCP, CD4-FITC, CD4-e450, CD4-PerCP, CD25-APC, IFNγ-APC, IL-2-PE, TNFa-PE-Cy7, CD11b-PE-Cy7, Ly6G-FITC, Ly6C-PerCP-Cy5.5, Gr1-PE-Cy7, and MHCII-EF450, PDGFRβ-APC, CD31-PE, CD45-BV510, and EpCam-BV605 were purchased from Thermo Fisher Scientific (Lafayette, CO) or BD Biosciences (San Jose, CA). CD8-PE-TxRD was purchased from Invitrogen (Carlsbad, CA). SIY, SIINFEKL, and β-galactosidase peptides were obtained from Integrated DNA Technologies (Coralville, Iowa). β-galactosidase (βgal) for the ICPMYARV peptide and SIINFEKL peptide tetramers were obtained from the NIH Tetramer core facility (Atlanta, GA). Antibodies to CD31 (Thermo Fisher Scientific), αSMA (Sigma-Aldrich, St Louis, MO), PDGFRα (Thermo Fisher Scientific), Vimentin (Cell Signaling Technology, MA), Ki67 (Abcam), Cleaved Caspase 3 (Cell Signaling Technology) were used for immunofluorescent staining and hypoxyprobe Omni Kit (Burlington, MA) was used for hypoxia staining. Anti-PD1 antibody clone RPM1-14 (BioXcell, West Lebanon, NH) was used where indicated for treatment studies.

Gunderson A.J., Yamazaki T., McCarty K., Phillips M., Alice A., Bambina S., Zebertavage L., Friedman D., Cottam B., Newell P., Gough M.J., Crittenden M.R., Van der Veken P, & Young K.H. (2019). Blockade of fibroblast activation protein in combination with radiation treatment in murine models of pancreatic adenocarcinoma. PLoS ONE, 14(2), e0211117.

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Isolation and Characterization of Mesoangioblasts

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Mesoangioblasts (MABs) were isolated from Gastrocnemius, Tibialis anterior, Quadriceps and Biceps explant culture, by Fluorescence Activated Cell Sorter (FACS; BD FACS ARIA III) for alkaline phosphatase (ALP) + cells (R&D Systems-Biotechne, Minneapolis, MN, USA) from 5 week-old mice as previously described [94 (link)]. Flow cytometry analysis of the ALP+ fraction was carried out in 3 WT and 7 Frzb^−/− samples. Protein tyrosine phosphatase receptor type C (CD45), platelet and endothelial cell adhesion molecule 1 (CD31) and platelet derived growth factor receptor alpha (PDGFRα or CD140α; Thermo Fisher Scientific) cell surface proteins presence were analysed by flow cytometry (FACs; BD Canto AIG) and analysed by BD FACSDiva software.

Casas-Fraile L., Cornelis F.M., Costamagna D., Rico A., Duelen R., Sampaolesi M.M., López de Munain A., Lories R.J, & Sáenz A. (2020). Frizzled related protein deficiency impairs muscle strength, gait and calpain 3 levels. Orphanet Journal of Rare Diseases, 15, 119.

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Proximity Ligation Assay for Protein Interactions

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PLA was performed with the Duolink in situ orange starter kit mouse/rabbit (Sigma-Aldrich Corp.) according to the manufacturer's instructions for custom solutions. Briefly, E13.5 embryos were fixed in 4% PFA in PBS and infiltrated with 30% sucrose in PBS before being mounted in OCT compound (Sakura Finetek USA, Inc.). Sections (8 µm) deposited on glass slides were fixed in 4% PFA in PBS with 0.1% Triton X-100 for 10 min and washed in PBS with 0.1% Triton X-100. Sections were blocked for 1 h in 5% normal donkey serum in PBS and incubated overnight at 4°C in primary antibodies diluted in 1% normal donkey serum in PBS. The following antibodies were used for PLA: PDGFRα (1:100; Thermo Fisher Scientific) and PDGFRβ (1:25; 28; Becton Dickinson). Sections were washed in PBS prior to incubation in PLA probes diluted in 1% normal donkey serum in PBS. Sections were photographed using an ORCA-Flash4.0 LT digital camera (Hamamatsu Photonics K.K.) fitted onto an AxioObserver.Z1 fluorescence microscope (Carl Zeiss, Inc.). Extended depth of focus was applied to z-stacks using Zen software (Carl Zeiss, Inc.) to generate images with the maximum depth of field. The number of puncta per 10 (mesenchyme) and five (epithelium) 50-µm² areas were counted in two independent experiments, and statistical analyses were performed with Prism 6 (GraphPad Software, Inc.) using a two-tailed unpaired t-test.

Fantauzzo K.A, & Soriano P. (2016). PDGFRβ regulates craniofacial development through homodimers and functional heterodimers with PDGFRα. Genes & Development, 30(21), 2443-2458.

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Kinase Activity Assay Protocol

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Myelin basic protein from bovine, DL-isocitric acid trisodium salt hydrate, β-nicotinamide adenine dinucleotide phosphate hydrate (NADP⁺), β-nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt hydrate (NADPH), α-ketoglutaric acid sodium salt, diaphorase from Clostridium kluyveri, resazurin sodium salt, and ATP disodium salt hydrate were purchased from Sigma-Aldrich. Ketoglutaric acid sodium salt, α-[1-¹⁴C], 50μCi (1.85MBq) and glucose, D-[U-¹⁴C]- were from PerkinElmer. Glutamine L-[5-¹⁴C] and isocitric acid[³H(G)] were from ARC. Inhibitors including dovitinib (TKI-258, CHIR-258), imatinib, AG-490, quizartinib, PP2, erlotinib, ruxolitinib, dasatinib and AG-120 were purchased from Selleckchem. Recombinant proteins including FLT3, SRC, PDGFRα, PDGFRβ, EGFR, MET, KIT, JAK2, FGFR3 and ABL1 were purchased from Thermo Fisher.

Chen D., Xia S., Wang M., Lin R., Li Y., Mao H., Aguiar M., Famulare C.A., Shih A.H., Brennan C.W., Gao X., Pan Y., Liu S., Fan J., Jin L., Song L., Zhou A., Mukherjee J., Pieper R.O., Mishra A., Peng J., Arellano M., Blum W.G., Lonial S., Boggon T.J., Levine R.L, & Chen J. (2019). Mutant and wild-type isocitrate dehydrogenase 1 share enhancing mechanisms involving distinct tyrosine kinase cascades in cancer. Cancer discovery, 9(6), 756-777.

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Multi-Marker Flow Cytometry Protocol

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FITC- and APC-CD19, CD11b, GR-1, FITC-TER119, FITC-CD44, PE-Dll1, Dll4, APC-c-kit, CD11c, B220, ST2, PE/Cy7-CD4, APC/Cy7-Thy1.2 and CD19 mAbs were all purchased from BioLegend (San Diego, CA). PE-CD19, PerCP/Cy5.5-CD25, APC-CD8, DX5, PDGFRα and PE/Cy7-CD45 mAbs were purchased from Thermo Fisher Scientific (Tokyo, Japan). Anti-NotchLs mAb, HRJ1-5, was established by immunization of Armenian hamsters with a recombinant rat Jagged1-human Fc chimeric protein (34) (R and D Systems Inc, Minneapolis, MN). Expression of NotchLs assessed by HRJ1-5 was detected by biotinylated goat anti-hamster IgG Ab (Thermo Fisher Scientific) and PE-conjugated streptavidin (BD Bioscience). Intracellular staining of HA-labeled Dll1 or Dll4 was detected by rabbit anti-HA mAb (Cell Signaling Technology) and DyLight649-conjugated donkey anti-rabbit IgG Ab (BioLegend). Expression of cell surface markers was analyzed by flow cytometry with a FACSVerse (BD Biosciences).

Hirano K.I., Suganami A., Tamura Y., Yagita H., Habu S., Kitagawa M., Sato T, & Hozumi K. (2020). Delta-like 1 and Delta-like 4 differently require their extracellular domains for triggering Notch signaling in mice. eLife, 9, e50979.

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Immunolabeling of Oligodendrocyte Progenitor Cells

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All steps were performed at ambient temperature. Cells were fixed with
4% paraformaldehyde in PBS for 5 minutes, permeabilized with 0.1% Triton X-100
containing PBS (except for PDGFRα and O4), blocked in 3% goat serum and
incubated with appropriate primary and secondary antibodies (Alexa Fluor
conjugated; Life Technologies). Nuclei were counterstained with DAPI (Sigma) and
coverslips were mounted on slides with FluorSave (Merck). Fields were selected
based upon uniform DAPI staining and imaged in three channels. Antibodies; OLIG2
(1:200, Millipore), PDGFRα (1:500, Cell Signalling), O4 (1:500,
R&D Systems), MBP (1:50, Abcam), GFAP (1:500, DAKO), GFAP (1:500, Cy3
conjugated; Sigma), CASPR (1:1000, Abcam), Sox2 (1:1000, Abcam), Oct3/4 (1:250,
Santa Cruz), Nanog (1:100, R&D Systems), TRA1-60 (1:100, StemGent), Human
Nuclei (1:300, Millipore), Human GFAP (1:1000, Biolegend), Neurofilament-H
(1:10,000, Biolegend).
Proportion of mitotic cells was carried out by labelling OPCs with
10μM ethynyl deoxy-uridine (EdU, Life Technologies) on day 3 for 24
hours. Media was replaced completely the next day and cells cultured for an
additional 48 hours in the absence of EdU before fixation using 4% PFA (Sigma).
For detection, cells were permeabilized using 0.1% saponin (Sigma) for 2 hours
and detected using the EdU detection kit (Life Technologies) according to
manufacturer’s instructions.

Vasistha N.A., Johnstone M., Barton S.K., Mayerl S.E., Thangaraj Selvaraj B., Thomson P.A., Dando O., Grünewald E., Alloza C., Bastin M.E., Livesey M.R., Economides K., Magnani D., Makedonopolou P., Burr K., Story D.J., Blackwood D.H., Wyllie D.J., McIntosh A.M., Millar J.K., ffrench-Constant C., Hardingham G.E., Lawrie S.M, & Chandran S. (2019). Familial t(1;11) translocation is associated with disruption of white matter structural integrity and oligodendrocyte–myelin dysfunction. Molecular Psychiatry, 24(11), 1641-1654.

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Immunocytochemical Characterization of Cultured Cells

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All steps were performed at ambient temperature. Cells were fixed with 4% paraformaldehyde in PBS for 5 min, permeabilized with 0.1% Triton X-100 containing PBS (except for PDGFRα and O4), blocked in 3% goat serum, and incubated with appropriate primary and secondary antibodies (Alexa Fluor conjugated; Life Technologies). Nuclei were counterstained with DAPI (Sigma) and coverslips were mounted on slides with FluorSave (Merck). Fields were selected based upon uniform DAPI staining and imaged in three channels. Antibodies; OLIG2 (1:200, Millipore), PDGFRα (1:500, Cell Signaling), O4 (1:500, R&D Systems), MBP (1:50, Abcam), GFAP (1:500, DAKO), GFAP (1:500, Cy3 conjugated; Sigma), CASPR (1:1000, Abcam), Sox2 (1:1000, Abcam), Oct3/4 (1:250, Santa Cruz), Nanog (1:100, R&D Systems), TRA1-60 (1:100, StemGent), hNu (1:300, Millipore), Human GFAP (1:1000, Biolegend), Neurofilament-H (1:10,000, Biolegend).
Proportion of mitotic cells was studied by labeling OPCs with 10 μM ethynyl deoxyuridine (EdU, Life Technologies) on day 3 for 24 h. Media was replaced completely the next day and cells cultured for an additional 48 h in the absence of EdU before fixation using 4% PFA (Sigma). For detection, cells were permeabilized using 0.1% saponin (Sigma) for 2 h and detected using the EdU detection kit (Life Technologies) according to manufacturer’s instructions.

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Transfection of ECs with siRNAs

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ECs at 50% confluence were transfected with non-targeting control siRNA (Qiagen, 1027280) or siRNAs targeting Snail (Life Technologies, s13185), Erg-1 (Life Technologies, s194569), NF-κB (Cell Signaling, 6261), PDGFR-α (Life Technologies, s10234), or PDGFR-β (Life Technologies, s10242) using Lipofectamine 2000 (Life Technologies, 11668-019) in serum-free Opti-MEM medium (GIBCO, 31985-070) for 12 h, followed by recovery with serum-supplemented medium for 24 h.

Liu T., Ma W., Xu H., Huang M., Zhang D., He Z., Zhang L., Brem S., O’Rourke D.M., Gong Y., Mou Y., Zhang Z, & Fan Y. (2018). PDGF-mediated mesenchymal transformation renders endothelial resistance to anti-VEGF treatment in glioblastoma. Nature Communications, 9, 3439.

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Differentiation Profiling of Cultured Cells

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Cultured cells were harvested at day (D)1.5, D2.5, D3.5 and D4.5 during the differentiation, stained with combinations of allophycocyanin (APC)‐conjugated anti‐Flk1 mAb (AVAS12) (Kataoka et al., 1997 ) and phycoerythrin (PE)‐conjugated anti‐platelet‐derived growth factor receptor α (PDGFRα) mAb (12‐1401‐81, eBioscience) and then subjected to analysis with a FACS Aria (Becton Dickinson) or CytoFLEX S (Beckman Coulter).

Minakawa T., Matoba T., Ishidate F., Fujiwara T.K., Takehana S., Tabata Y, & Yamashita J.K. (2021). Extracellular vesicles synchronize cellular phenotypes of differentiating cells. Journal of Extracellular Vesicles, 10(11), e12147.

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FACS Analysis of Proliferative Hair Follicle Cells

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FACSVerse^TM (BD Biosciences, Franklin Lakes, NJ, USA) was utilized to identify the level of proliferative EGFP-positive (EGFP⁺) cells as well as that of mesenchymal stem cells among proliferative EGFP⁺ cells. Both freshly isolated and proliferative hair follicle cells were stained with anti-platelet-derived growth factor α (PDGFRα) and anti-stem cell antigen-1 (Sca-1) (e-Bioscience, Santa Clara, CA, USA) for 30 minutes on ice, and then compared.

Urano-Morisawa E., Takami M., Suzawa T., Matsumoto A., Osumi N., Baba K, & Kamijo R. (2017). Induction of osteoblastic differentiation of neural crest-derived stem cells from hair follicles. PLoS ONE, 12(4), e0174940.

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