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Flow cytometric analysis of Chromogranin A expression in Human neuroblastoma cells. Cells from the Human SH-SY5Y (Neuroblastoma; ATCC® CRL-2266™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and then washed, permeabilized with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human Chromogranin A antibody (Cat. No. 569823/569898; solid line histogram). The fluorescence histogram showing Chromogranin A expression (or Ig Isotype control staining) was derived from gated events with the forward and side-light scatter characteristics of intact SH-SY5Y cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BD Horizon™ BV421 Mouse Anti-Human Chromogranin A
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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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Chromogranin A (CGA) is a member of the granin family of regulated secretory proteins that are found in secretory granules in endocrine and neuroendocrine cells and released in response to extracellular stimulation. Intracellularly, granins are important for targeting peptide hormones and neurotransmitters by their ability to aggregate in the low pH, high calcium environment of the trans-Golgi network. Extracellularly, peptides formed from proteolytic processing of granins regulate hormone secretion. CGA is a prohormone that can be cleaved into several biologically active peptides, such as pancreastatin, β-granin, vasostatin, catestatin, and parastatin. β-granin is an N-terminal fragment of CGA, while pancreastatin and catestatin are processed from the central region of CGA. Cells of the adrenal medulla, anterior pituitary, cerebral cortex as well as beta cells of the pancreas and a variety of tumor cell lines express CGA. The expression of CGA can be used to monitor the pancreatic differentiation of pluripotent stem cells.
Development References (7)
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D'Amour KA, Bang AG, Eliazer S, et al . Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol. 2006; 24(12):1481-1483. (Biology). View Reference
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Hendy GN, Bevan S, Mattei MG, Mouland AJ. Chromogranin A. Clin Invest Med. 1995; 18(1):47-65. (Biology). View Reference
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Kroon E, Martinson LA, Kadoya K, Bang AG, et al. Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat Biotechnol. 2008; 26(4):443-452. (Biology). View Reference
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Loh YP, Cheng Y, Mahata SK, Corti A, Tota B. Chromogranin A and derived peptides in health and disease. J Mol Neurosci. 2012; 48(2):347-356. (Biology). View Reference
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Mouland AJ, Bevan S, White JH, Hendy GN. Human chromogranin A gene. Molecular cloning, structural analysis, and neuroendocrine cell-specific expression. J Biol Chem. 1994; 269(9):6918-6926. (Biology). View Reference
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Schulz TC, Young HY, Agulnick AD et al. A scalable system for production of functional pancreatic progenitors from human embryonic stem cells. PLoS ONE. 7(5)(Biology). View Reference
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Taylor CV, Taupenot L, Mahata SK. Formation of the catecholamine release-inhibitory peptide catestatin from chromogranin A. Determination of proteolytic cleavage sites in hormone storage granules. Clin Invest Med. 2000; 275(30):22905-22915. (Biology). View Reference
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