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Flow cytometric analysis of CD140a (PDGFRα) expression on Human MG-63 cells. Cells from the Human MG-63 (Osteosarcoma, ATCC CRL-1427) cell line were stained with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dotted line histogram) or with BD Horizon™ BV421 Mouse Anti-Human CD140a (PDGFRα) antibody (Cat. No. 568238/568239; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The histogram showing CD140a (PDGFRα) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) MG-63 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Horizon™ BV421 Mouse Anti-Human CD140a (PDGFRα)
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BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
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The 16A1 monoclonal antibody specifically recognizes human CD140a which is also known as human platelet derived growth factor receptor α (PDGFRα). CD140a (PDGFRα) is an ~170 kDa single pass type I transmembrane glycoprotein that is encoded by PDGFRA (platelet derived growth factor receptor alpha) which belongs to the receptor tyrosine kinase (RTK) class III family. This receptor is comprised of an N-terminal extracellular domain with five IgC2-like domains, and a transmembrane sequence followed by a cytoplasmic region with a split tyrosine kinase domain. CD140a (PDGFRα) is expressed on a variety of cell types including platelets, monocytes, fibroblasts, endothelial cells, smooth muscle cells, glial cells, chondrocytes as well as numerous tumor cell types. PDGFRα dimerizes and autophosphorylates when binding to homodimeric or heterodimeric platelet-derived growth factors (PDGF) comprised of disulfide-bonded A, B, or C (but not D) isoforms, ie, PDGF-AA, PDGF-AB, PDGF-BB, and PDGF-CC. PDGFRα can also heterodimerize with platelet derived growth factor receptor β (PDGFRβ, also known as CD140b) and autophosphorylate upon binding to PDGF-AB, PDGF-BB, and possibly PDGF-CC and PDGF-DD. Homodimerized CD140b (PDGFRβ) is activated by PDGF-BB and PDGF-DD. PDGF ligands stimulate the growth, proliferation, migration, and survival of PDGF Receptor-positive normal cells as well as tumor cells. CD140a (PDGFRα) plays key roles in multiple processes including embryonic development, wound healing, and the regulation of platelet activation.
研发参考 (4)
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Demoulin JB, Essaghir A. PDGF receptor signaling networks in normal and cancer cells. Cytokine Growth Factor Rev. 2014; 25(3):273-283. (Biology). 查看参考
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Hara Y, Yamashita T, Oishi N, et al. TSU-68 ameliorates hepatocellular carcinoma growth by inhibiting microenvironmental platelet-derived growth factor signaling.. Anticancer Res. 2015; 35(3):1423-31. (Clone-specific: Flow cytometry). 查看参考
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Hart CE, Bowen-Pope DF. CD140a and b (PDGRα and β) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:739-741.
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Miyazaki S, Sugawara H, Tamura T. Cross-lineage (Blind Panel) study and human leucocyte differentiation antigen database. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:3-20.
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