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      BV480 Mouse Anti-Ki-67
      BV480 Mouse Anti-Ki-67
      Two-color flow cytometric analysis of Ki-67 expression by proliferating MOLT-4 and human peripheral blood mononuclear cells (PBMC). Proliferating MOLT-4 cells and predominantly noncycling PBMC were fixed and permeabilized separately with 70% ice cold ethanol, washed, and stained with BD Horizon™ BV480 Mouse Anti-Ki-67 antibody (Cat. No. 566109/566172). The cells were then counterstained with BD Via-Probe™ Cell Viability Solution [Cat. No. 555815/555816; contains 7-Amino-Actinomycin D (7-AAD)] to stain DNA. Two-color flow cytometric dot plots showing the correlated expression patterns of 7-AAD staining versus Ki-67 were derived from gated events with the forward and side light-scatter characteristics of intact MOLT-4 cells (Left Plot) or PBMC (Right Plot). Flow cytometric analysis was performed using a BD LSR Fortessa™ Cell Analyzer System.
      BV480 Mouse Anti-Ki-67
      Two-color flow cytometric analysis of Ki-67 expression by proliferating MOLT-4 and human peripheral blood mononuclear cells (PBMC). Proliferating MOLT-4 cells and predominantly noncycling PBMC were fixed and permeabilized separately with 70% ice cold ethanol, washed, and stained with BD Horizon™ BV480 Mouse Anti-Ki-67 antibody (Cat. No. 566109/566172). The cells were then counterstained with BD Via-Probe™ Cell Viability Solution [Cat. No. 555815/555816; contains 7-Amino-Actinomycin D (7-AAD)] to stain DNA. Two-color flow cytometric dot plots showing the correlated expression patterns of 7-AAD staining versus Ki-67 were derived from gated events with the forward and side light-scatter characteristics of intact MOLT-4 cells (Left Plot) or PBMC (Right Plot). Flow cytometric analysis was performed using a BD LSR Fortessa™ Cell Analyzer System.
      Product Details
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      BD Horizon™
      MKI67; Antigen identified by monoclonal antibody Ki-67; KIA
      Human (QC Testing), Mouse (Tested in Development), Rat, Rhesus (Reported)
      Mouse IgG1, κ
      Human Ki-67
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2739511
      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 BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 were removed.
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      Recommended Assay Procedures

      Immunofluorescent Staining and Flow Cytometric Analysis: The B56 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify proliferating cells. The BD Horizon™ BV480 Anti-Ki-67 antibody (Cat. No. 566109/566172) is suitable for these experiments. For specific methodology, please visit our web site, www.bdbiosciences.com/us/s/resources, and go to the "Flow Cytometry Staining Protocol for Detection of Ki-67" under "Multicolor Flow Cytometry".

      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. 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).
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. 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.
      5. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      Antibody Details
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      B56

      The B56 monoclonal antibody specifically binds to the Ki-67 antigen that is expressed in the nucleus of cycling cells (G1, S, G2, M cell cycle phases). During the G0 phase, the antigen cannot be detected. During interphase of the cell cycle, it is associated with nucleolar components, and it is on the surface of the chromosomes during M phase. Ki-67 is a large protein having 2 alternatively spliced isoforms, an N-terminal forkhead-associated domain, a C-terminal domain that binds to heterochromatin proteins, and multiple phosphorylation sites, the functions of which are still unclear. Because of the strict association of Ki-67 expression with cell proliferation, anti-Ki-67 antibodies are useful for the identification, quantification, and monitoring of growing cell populations.

      The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

      Format Details
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      BV480
      The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV480
      Violet 405 nm
      440 nm
      479 nm
      Citations & References
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      View product citations for antibody "566109" on CiteAb

      Development References (14)

      1. Benson MJ, Elgueta R, Schpero W, et al. Distinction of the memory B cell response to cognate antigen versus bystander inflammatory signals. J Exp Med. 2009; 206(9):2013-2025. (Clone-specific: Flow cytometry). View Reference
      2. Bigley V, Haniffa M, Doulatov S, et al. The human syndrome of dendritic cell, monocyte, B and NK lymphoid deficiency. J Exp Med. 2011; 208(2):227-234. (Clone-specific: Flow cytometry). View Reference
      3. Bruno S, Crissman HA, Bauer KD, Darzynkiewicz Z. Changes in cell nuclei during S phase: progressive chromatin condensation and altered expression of the proliferation-associated nuclear proteins Ki-67, cyclin (PCNA), p105, and p34. Exp Cell Res. 1991; 196(1):99-106. (Biology: Flow cytometry). View Reference
      4. Bruno S, Darzynkiewicz Z. Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells. Cell Prolif. 1992; 25(1):31-40. (Biology: Flow cytometry). View Reference
      5. Kill IR. Localisation of the Ki-67 antigen within the nucleolus: evidence for a fibrillarin-deficient region of the dense fibrillar component. J Cell Sci. 1996; 109(6):1253-1263. (Biology). View Reference
      6. Kouro T, Medina KL, Oritani K, Kincade PW. Characteristics of early murine B-lymphocyte precursors and their direct sensitivity to negative regulators. Blood. 2001; 97(9):2708-2715. (Clone-specific: Flow cytometry). View Reference
      7. Kubbutat MH, Key G, Duchrow M, Schluter C, Flad HD, Gerdes J. Epitope analysis of antibodies recognising the cell proliferation associated nuclear antigen previously defined by the antibody Ki-67 (Ki-67 protein). J Clin Pathol. 1994; 47(6):524-528. (Biology). View Reference
      8. Picker LJ, Hagen SI, Lum R, et al. Insufficient production and tissue delivery of CD4+ memory T cells in rapidly progressive simian immunodeficiency virus infection. J Exp Med. 2004; 200(10):1299-1314. (Clone-specific: Flow cytometry). View Reference
      9. Pitcher CJ, Hagen SI, Walker JM, et al. Development and homeostasis of T cell memory in rhesus macaque. J Immunol. 2002; 168(1):29-43. (Clone-specific: Flow cytometry). View Reference
      10. Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown.. J Cell Physiol. 2000; 182(3):311-22. (Biology). View Reference
      11. Shi SR, Key ME, Kalra KL. Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. J Histochem Cytochem. 1991; 39(6):741-748. (Biology). View Reference
      12. Spargo LDJ, Cleland LG, Cockshell MP, Mayrhofer Graham. Recruitment and proliferation of CD4+ T cells in synovium following adoptive transfer of adjuvant-induced arthritis. Int Immunol. 2006; 18(6):897-910. (Clone-specific: Flow cytometry, Immunofluorescence).
      13. Starborg M, Gell K, Brundell E, Höög C. The murine Ki-67 cell proliferation antigen accumulates in the nucleolar and heterochromatic regions of interphase cells and at the periphery of the mitotic chromosomes in a process essential for cell cycle progression. J Cell Sci. 1996; 109(1):143-153. (Biology). View Reference
      14. Valenti LM, Mathieu J, Chancerelle Y, et al. High levels of endogenous nitric oxide produced after burn injury in rats arrest activated T lymphocytes in the first G1 phase of the cell cycle and then induce their apoptosis. Exp Cell Res. 2005; 306(1):150-167. (Clone-specific: Flow cytometry). View Reference
      View All (14) View Less

       

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