Skip to main content Skip to navigation
V450 Mouse anti-Ki-67
V450 Mouse anti-Ki-67

Flow cytometric analysis of Ki-67 expression by proliferating Jurkat and noncycling human peripheral blood mononuclear cells (PBMC). Human Jurkat and PBMC were fixed and permeabilized with 70% ice cold ethanol, washed, and stained  with BD Horizon™ V450 Mouse anti-Ki-67 antibody (Cat. No. 561281) according to the BD Biosciences support protocol, Flow Cytometry Staining Protocol for Detection of Ki-67. The cells were then RNase A (Sigma Cat. No. R-5500) treated and counterstained with Propidium Iodide Staining Solution (Cat. No. 556463) to stain double-stranded DNA. A two-color flow cytometric dot plot showing the correlated expression patterns of Propidium Iodide (DNA) staining versus Ki-67 was derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells (Left Panel) or PBMC (Right Panel). Flow cytometry was performed using a BD LSR™ II Flow Cytometer System.

Flow cytometric analysis of Ki-67 expression by proliferating Jurkat and noncycling human peripheral blood mononuclear cells (PBMC). Human Jurkat and PBMC were fixed and permeabilized with 70% ice cold ethanol, washed, and stained  with BD Horizon™ V450 Mouse anti-Ki-67 antibody (Cat. No. 561281) according to the BD Biosciences support protocol, Flow Cytometry Staining Protocol for Detection of Ki-67. The cells were then RNase A (Sigma Cat. No. R-5500) treated and counterstained with Propidium Iodide Staining Solution (Cat. No. 556463) to stain double-stranded DNA. A two-color flow cytometric dot plot showing the correlated expression patterns of Propidium Iodide (DNA) staining versus Ki-67 was derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells (Left Panel) or PBMC (Right Panel). Flow cytometry was performed using a BD LSR™ II Flow Cytometer System.

製品詳細
Down Arrow Up Arrow


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_10613816
Aqueous buffered solution containing protein stabilizer 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™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.

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. 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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  4. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561281 Rev. 2
抗体の詳細
Down Arrow Up Arrow
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 is conjugated to BD Horizon V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon V450 can be used in place of Pacific Blue™ conjugates.

561281 Rev. 2
フォーマットの詳細
Down Arrow Up Arrow
V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
V450
Violet 405 nm
405 nm
450 nm
561281 Rev.2
引用&参考文献
Down Arrow Up Arrow

Development References (11)

  1. 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
  2. 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
  3. Byeon IJ, Li H, Song H, Gronenborn AM, Tsai MD. Sequential phosphorylation and multisite interactions characterize specific target recognition by the FHA domain of Ki67.. Nat Struct Mol Biol. 2005; 12(11):987-93. (Biology). View Reference
  4. Ho DWY, Fan ST, To J, et al. Selective plasma filtration for treatment of fulminant hepatic failure induced by D-galactosamine in a pig model. Gut. 2002; 50:869-876. (Clone-specific).
  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. 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
  8. Scholzen T, Endl E, Wohlenberg C, et al. The Ki-67 protein interacts with members of the heterochromatin protein 1 (HP1) family: a potential role in the regulation of higher-order chromatin structure.. J Pathol. 2002; 196(2):135-44. (Biology). View Reference
  9. 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
  10. 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).
  11. 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
すべて表示する (11) 表示項目を減らす
561281 Rev. 2

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.