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      Purified Mouse Anti-Human Ki-67
      Purified Mouse Anti-Human Ki-67
      Flow cytometric analysis of Ki-67 expression in proliferating Human MOLT-4 cells. Proliferating cells from the Human MOLT-4 (T lymphoblastic leukemia, ATCC® CRL-1582™) cell line were permeabilized and fixed with 70% ice-cold ethanol. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], labeled with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; dashed line histogram) or Purified Mouse Anti-Ki-67 antibody (Cat. No. 571599; solid line histogram) at 0.25 µg/test. The cells were then secondarily stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) [Cat. No. 550589]. Histograms showing Ki-67 level (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Purified Mouse Anti-Human Ki-67
      Immunofluorescent staining of Ki-67 expressed in Human HeLa cells. Proliferating cells from the HeLa (Cervical  Adenocarcinoma, ATCC® CRM-CCL-2™) cell line were cultured, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). Cells were labeled with Purified Mouse Anti-Ki-67 antibody (Cat. No. 571599), washed, and secondarily stained with Alexa Fluor™ 488 Goat Anti-Mouse Ig (pseudocolored green) and counter-stained with BD Pharmingen™ DRAQ7™ (Cat. No. 564904, pseudocolored red). Images were captured using a standard epifluorescence microscope with a 20x objective. Data shown on this Technical Data Sheet are not lot specific.
      Purified Mouse Anti-Human Ki-67 Purified Mouse Anti-Human Ki-67
      Flow cytometric analysis of Ki-67 expression in proliferating Human MOLT-4 cells. Proliferating cells from the Human MOLT-4 (T lymphoblastic leukemia, ATCC® CRL-1582™) cell line were permeabilized and fixed with 70% ice-cold ethanol. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], labeled with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; dashed line histogram) or Purified Mouse Anti-Ki-67 antibody (Cat. No. 571599; solid line histogram) at 0.25 µg/test. The cells were then secondarily stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) [Cat. No. 550589]. Histograms showing Ki-67 level (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Immunofluorescent staining of Ki-67 expressed in Human HeLa cells. Proliferating cells from the HeLa (Cervical  Adenocarcinoma, ATCC® CRM-CCL-2™) cell line were cultured, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). Cells were labeled with Purified Mouse Anti-Ki-67 antibody (Cat. No. 571599), washed, and secondarily stained with Alexa Fluor™ 488 Goat Anti-Mouse Ig (pseudocolored green) and counter-stained with BD Pharmingen™ DRAQ7™ (Cat. No. 564904, pseudocolored red). Images were captured using a standard epifluorescence microscope with a 20x objective. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      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), Immunofluorescence (Tested During Development)
      0.5 mg/ml
      4288
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
      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. For U.S. patents that may apply, see bd.com/patents.
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      571599 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.

      571599 Rev. 1
      Format Details
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      Purified
      Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
      Purified
      571599 Rev.1
      Citations & References
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      View product citations for antibody "571599" 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
      571599 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.