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BV510 Mouse Anti-Ki-67
BV510 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™ BV510 Mouse Anti-Ki-67 antibody (Cat. No. 563462) according to the BD Biosciences support protocol, \"Flow Cytometry Staining Protocol for Detection of Ki-67.\" 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 Panel) or PBMC (Right Panel). Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometer System.
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™ BV510 Mouse Anti-Ki-67 antibody (Cat. No. 563462) according to the BD Biosciences support protocol, \"Flow Cytometry Staining Protocol for Detection of Ki-67.\" 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 Panel) or PBMC (Right Panel). Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometer 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_2738221
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™ BV510 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV510 were 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. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Brilliant Violet™ 510 is a trademark of Sirigen.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563462 Rev. 2
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 BV510 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 405-nm and Em Max at 510-nm, BD Horizon BV510 can be excited by the violet laser and detected in the BD Horizon V500 (525/50-nm) filter set. BD Horizon BV510 conjugates are useful for the detection of dim markers off the violet laser.

563462 Rev. 2
Format Details
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BV510
The BD Horizon Brilliant Violet™ 510 (BV510) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye with an excitation maximum (Ex Max) at 327-nm / 405-nm and an emission maximum (Em Max) at 512-nm. BV510, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 510-nm (e.g., a 525/50 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.
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BV510
Violet 405 nm
327 nm, 405 nm
512 nm
563462 Rev.2
Citations & References
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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
View All (11) View Less
563462 Rev. 2

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