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    BV421 Mouse Anti-Glucagon
    BV421 Mouse Anti-Glucagon
    Flow cytometric analysis of Glucagon expression in a mouse pancreatic α cell line. Alpha TC1-6 cells (ATCC CRL-2934) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed and then stained with either BD Horizon™ BV421 Mouse IgG1, k Isotype Control (Cat. No. 562438, dotted-line histogram) or BD Horizon BV421 Mouse Anti-Glucagon (Cat. No. 565891, solid-line histogram). The fluorescence histogram was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
    BV421 Mouse Anti-Glucagon
    Immunohistofluorescent staining of Glucagon in human, mouse, and rat islets of Langerhans. Following antigen retrieval with BD Pharmingen™ Retrievagen A Buffer (Cat. No. 550524), sections from  formalin-fixed, paraffin-embedded human (top row), mouse (middle row), and rat (bottom row) pancreata were blocked using an Avidin/ Biotin Blocking Kit (Vector Laboratories, Cat. No. SP-2001) as recommended by the manufacturer. LEFT PANEL: These sections were then stained with BD Horizon BV421 Mouse Anti-Glucagon (Cat. No. 565891, pseudocolored red), and cell nuclei were counterstained with BD Pharmingen™ DRAQ5™ (Cat. No. 564902 pseudo-colored green). RIGHT PANEL: These sections were then stained with BD Horizon BV421 Mouse Anti-Glucagon (Cat. No. 565891, pseudocolored red) and Alexa Fluor® 647 Mouse Anti-Insulin (Cat. No.565689, pseudocolored green). The cell nuclei were counterstained with BD Pharmingen™ 7-AAD (Cat. No. 559925, pseudocolored blue, top and middle rows only). The typical localization of the α cells is seen: distributed throughout human islets and at the periphery in rodent islets. The photographs were performed on a standard  epifluorescence microscope. Original magnification, 20x.
    BV421 Mouse Anti-Glucagon BV421 Mouse Anti-Glucagon
    Flow cytometric analysis of Glucagon expression in a mouse pancreatic α cell line. Alpha TC1-6 cells (ATCC CRL-2934) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed and then stained with either BD Horizon™ BV421 Mouse IgG1, k Isotype Control (Cat. No. 562438, dotted-line histogram) or BD Horizon BV421 Mouse Anti-Glucagon (Cat. No. 565891, solid-line histogram). The fluorescence histogram was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
    Immunohistofluorescent staining of Glucagon in human, mouse, and rat islets of Langerhans. Following antigen retrieval with BD Pharmingen™ Retrievagen A Buffer (Cat. No. 550524), sections from  formalin-fixed, paraffin-embedded human (top row), mouse (middle row), and rat (bottom row) pancreata were blocked using an Avidin/ Biotin Blocking Kit (Vector Laboratories, Cat. No. SP-2001) as recommended by the manufacturer. LEFT PANEL: These sections were then stained with BD Horizon BV421 Mouse Anti-Glucagon (Cat. No. 565891, pseudocolored red), and cell nuclei were counterstained with BD Pharmingen™ DRAQ5™ (Cat. No. 564902 pseudo-colored green). RIGHT PANEL: These sections were then stained with BD Horizon BV421 Mouse Anti-Glucagon (Cat. No. 565891, pseudocolored red) and Alexa Fluor® 647 Mouse Anti-Insulin (Cat. No.565689, pseudocolored green). The cell nuclei were counterstained with BD Pharmingen™ 7-AAD (Cat. No. 559925, pseudocolored blue, top and middle rows only). The typical localization of the α cells is seen: distributed throughout human islets and at the periphery in rodent islets. The photographs were performed on a standard  epifluorescence microscope. Original magnification, 20x.
    Product Details
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    BD Horizon™
    GLP1; GLP2; GRPP; GCG
    Human (QC Testing), Mouse, Rat (Tested in Development)
    Mouse BALB/c IgG1, κ
    Human glucagon Recombinant Protein
    Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence, Immunohistochemistry-paraffin (Tested During Development)
    0.2 mg/ml
    AB_2739385
    Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
    RUO


    Preparation And Storage

    Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
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    Recommended Assay Procedures

    For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

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    Product Notices

    1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
    2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
    3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
    4. DRAQ5™ is a registered trademark of BioStatus Ltd.
    5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
    6. 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.
    7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
    9. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
    10. An isotype control should be used at the same concentration as the antibody of interest.
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    565891 Rev. 1
    Antibody Details
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    U16-850

    The U16-850 monoclonal antibody specifically binds to glucagon, a member of the secretin family of active peptides. Glucagon is an evolutionarily conserved peptide hormone that participates in the regulation of carbohydrate metabolism by counteracting the effects of insulin. Glucagon is produced by α cells in the islets of Langerhans of the pancreas. The CGC gene encodes the precursor molecule preproglucagon, which is cleaved to form proglucagon that is in turn cleaved to form at least four distinct peptides, including glucagon, in different tissues. Hypoglycemia causes the secretion of glucagon, which binds to the class B G-protein-coupled glucagon receptor that is mainly expressed in liver and kidney, causing reduced glycogenesis and glycolysis and increased glycogenolysis and gluconeogenesis. The expression of glucagon can be used to monitor the pancreatic differentiation of pluripotent stem cells.

    The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant Violet™ family of dyes.  With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter).  BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

    565891 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.
    altImg
    BV421
    Violet 405 nm
    407 nm
    423 nm
    565891 Rev.1
    Citations & References
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    Development References (6)

    1. Bowerman M, Michalski JP, Beauvais A, Murray LM, DeRepentigny Y, Kothary R. Defects in pancreatic development and glucose metabolism in SMN-depleted mice independent of canonical spinal muscular atrophy neuromuscular pathology.. Hum Mol Genet. 2014; 23(13):3432-44. (Biology). View Reference
    2. 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
    3. Jiang G, Zhang BB. Glucagon and regulation of glucose metabolism.. Am J Physiol Endocrinol Metab. 2003; 284(4):E671-8. (Biology). View Reference
    4. Kelly OG, Chan MY, Martinson LA, et al. Cell-surface markers for the isolation of pancreatic cell types derived from human embryonic stem cells. Nat Biotechnol. 2011; 29(8):750-756. (Biology). View Reference
    5. Thomsen J, Kristiansen K, Brunfeldt K, Sundby F. The amino acid sequence of human glucagon.. FEBS Lett. 1972; 21(3):315-319. (Biology). View Reference
    6. Witt S, Dietz H, Ziegler B, Keilacker H, Ziegler M. Production and use of monoclonal glucagon and insulin antibodies--reduction of pancreatic insulin in rats by treatment with complete Freund's adjuvant. Acta Histochem Suppl. 1988; 35:217-223. (Biology). View Reference
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    565891 Rev. 1

    Please refer to Support Documents for Quality Certificates


    Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


    Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

    For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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