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BV421 Mouse Anti-Human Chromogranin A
BV421 Mouse Anti-Human Chromogranin A
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.
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.
Product Details
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BD Horizon™
CGA; CgA; CHGA; Chromogranin-A; CMGA; SP-I
Human (QC Testing)
Mouse IgG1, κ
Human Chromogranin A Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl/test
1113
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

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).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. 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).
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  10. For U.S. patents that may apply, see bd.com/patents.
569898 Rev. 1
Antibody Details
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S21-537

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.

569898 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
569898 Rev.1
Citations & References
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Development References (7)

  1. 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
  2. Hendy GN, Bevan S, Mattei MG, Mouland AJ. Chromogranin A. Clin Invest Med. 1995; 18(1):47-65. (Biology). View Reference
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
View All (7) View Less
569898 Rev. 1

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