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RY610 Mouse Anti-Human Ki-67
RY610 Mouse Anti-Human Ki-67
Multiparameter flow cytometric analysis of Ki-67 expression in noncycling Human peripheral blood lymphocytes or proliferating Human MOLT-4 cells. Noncycling normal Human peripheral blood mononuclear cells containing lymphocytes (Top Plots) or proliferating cells from the Human MOLT-4 (T lymphoblastic leukemia, ATCC® CRL-1582™) cell line (Bottom Plots) were permeabilized and fixed with 70% ice-cold ethanol. In separate experiments, the normal or MOLT-4 cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], stained with either BD Horizon™ RY610 Mouse IgG1, κ Isotype Control (Cat. No. 571151; Left Plots) or BD Horizon™ RY610 Mouse Anti-Human Ki-67 antibody (Cat. No. 571266/571326; Right Plots) and counterstained with BD Pharmingen™ DAPI Solution (Cat. No. 564907) to stain DNA. Bivariate pseudocolor density plots showing the correlated expression of DNA (DAPI) versus Ki-67 levels (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes or MOLT-4 cells. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of Ki-67 expression in noncycling Human peripheral blood lymphocytes or proliferating Human MOLT-4 cells. Noncycling normal Human peripheral blood mononuclear cells containing lymphocytes (Top Plots) or proliferating cells from the Human MOLT-4 (T lymphoblastic leukemia, ATCC® CRL-1582™) cell line (Bottom Plots) were permeabilized and fixed with 70% ice-cold ethanol. In separate experiments, the normal or MOLT-4 cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], stained with either BD Horizon™ RY610 Mouse IgG1, κ Isotype Control (Cat. No. 571151; Left Plots) or BD Horizon™ RY610 Mouse Anti-Human Ki-67 antibody (Cat. No. 571266/571326; Right Plots) and counterstained with BD Pharmingen™ DAPI Solution (Cat. No. 564907) to stain DNA. Bivariate pseudocolor density plots showing the correlated expression of DNA (DAPI) versus Ki-67 levels (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes or MOLT-4 cells. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
Product Details
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BD Horizon™
Antigen KI-67; Antigen Ki67; KI-67; KI67; MKI67
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human L428 Hodgkin Lymphoma Cell Line Nuclei
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl/test
4288
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 unreacted dye was 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.

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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. 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.
  8. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  9. CF™ is a trademark of Biotium, Inc.
  10. For U.S. patents that may apply, see bd.com/patents.
571326 Rev. 1
Antibody Details
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Ki-67

The Ki-67 monoclonal antibody specifically recognizes the Ki-67 antigen (also known as KI67, KI-67). Ki-67 is a nonhistone nuclear protein that is encoded by MKI67 (Marker of Proliferation Ki-67). Ki-67 is associated with cellular proliferation and its expression increases as cells enter and progress through the G1, S, G2, and M cell cycle phases. Ki-67 is a large protein having 2 alternatively spliced isoforms. The Ki-67 protein contains an N-terminal forkhead-associated domain, a protein phosphatase 1-binding domain, a large central region of tandem repeats, and a C-terminal leucine/arginine-rich chromatin-binding domain. Ki-67 is required for cell proliferation and serves as useful marker for distinguishing and analyzing quiescent cells (Ki-67-negative cells within the G0 cell cycle phase) from cycling Ki-67-positive cells including cancer cells.

571326 Rev. 1
Format Details
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RY610
The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
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RY610
Yellow-Green 561 nm
557 nm
610 nm
571326 Rev.1
Citations & References
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View product citations for antibody "571326" on CiteAb

Development References (8)

  1. 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). View Reference
  2. 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
  3. Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H. Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67.. J Immunol. 1984; 133(4):1710-5. (Clone-specific: Immunocytochemistry). View Reference
  4. Gerdes J, Schwab U, Lemke H, Stein H. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation.. Int J Cancer. 1983; 31(1):13-20. (Immunogen: Immunohistochemistry). View Reference
  5. Kim KH, Sederstrom JM. Assaying Cell Cycle Status Using Flow Cytometry.. Curr Protoc Mol Biol. 2015; 111:28.6.1-28.6.11. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
  6. Palutke M, KuKuruga D, Tabaczka P. A flow cytometric method for measuring lymphocyte proliferation directly from tissue culture plates using Ki-67 and propidium iodide.. J Immunol Methods. 1987; 105(1):97-105. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
  7. Schwarting R, Gerdes J, Niehus J, Jaeschke L, Stein H. Determination of the growth fraction in cell suspensions by flow cytometry using the monoclonal antibody Ki-67.. J Immunol Methods. 1986; 90(1):65-70. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
  8. Sun X, Kaufman PD. Ki-67: more than a proliferation marker. Chromosoma. 2018; 127(2):175-186. (Biology). View Reference
View All (8) View Less
571326 Rev. 1

 

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