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      RY775 Mouse Anti-Ki-67
      RY775 Mouse Anti-Ki-67
      Multiparameter flow cytometric analysis of Ki-67 expression in noncycling Human peripheral blood lymphocytes or proliferating Human MOLT-4 cells.  Noncycling 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. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], stained with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plots) or BD Horizon™ RY775 Mouse Anti-Ki-67 antibody (Cat. No. 571398/571399; 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 PBMC (Top Plots) or MOLT-4 cells (Bottom Plots).Samples were acquired using the BD FACSymphony™ A5 SE Cell Analyzer System and were spectrally unmixed using FlowJo™ v10.10 Software.
      RY775 Mouse Anti-Ki-67
      Multiparameter flow cytometric analysis of Ki-67 expression in noncycling Human peripheral blood lymphocytes or proliferating Human MOLT-4 cells.  Noncycling 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. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], stained with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plots) or BD Horizon™ RY775 Mouse Anti-Ki-67 antibody (Cat. No. 571398/571399; 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 PBMC (Top Plots) or MOLT-4 cells (Bottom Plots).Samples were acquired using the BD FACSymphony™ A5 SE Cell Analyzer System and were spectrally unmixed using FlowJo™ v10.10 Software.
      商品详情
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      BD Horizon™
      MKI67; Antigen identified by monoclonal antibody Ki-67; KIA
      Human (QC Testing), Mouse (Tested in Development), Rat,Rhesus,Cynomolgus,Baboon,Dog,Chicken (Reported)
      Mouse IgG1, κ
      Human Ki-67
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl/test
      4288
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      准备和存储

      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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      推荐的实验流程

      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.

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      商品通知

      1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      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. An isotype control should be used at the same concentration as the antibody of interest.
      5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      8. 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.
      9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      10. 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.
      11. 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.
      12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
      13. For U.S. patents that may apply, see bd.com/patents.
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      571399 Rev. 1
      抗体详情
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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.

      571399 Rev. 1
      格式详情
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      RY775
      The BD Horizon RealYellow™ 775 (RY775) 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 775-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY775 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), RY775 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter).
      altImg
      RY775
      Yellow-Green 561 nm
      557 nm
      775 nm
      571399 Rev.1
      报价单和参考
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      View product citations for antibody "571399" on CiteAb

      研发参考 (13)

      1. Alitheen NB, McClure SJ, Yeap SK, Kristeen-Teo YW, Tan SW, McCullagh P. Establishment of an in vitro system representing the chicken gut-associated lymphoid tissue.. PLoS One. 2012; 7(11):e49188. (Clone-specific: Immunohistochemistry). 查看参考
      2. 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: Intracellular Staining/Flow Cytometry). 查看参考
      3. 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: Intracellular Staining/Flow Cytometry). 查看参考
      4. Buechler C, Semler M, Baker DA, et al. Subclinical Infection of Macaques and Baboons with A Baboon Simarterivirus.. Viruses. 2018; 10(12):701. (Clone-specific: Intracellular Staining/Flow Cytometry). 查看参考
      5. Kim KH, Sederstrom JM. Assaying Cell Cycle Status Using Flow Cytometry.. Curr Protoc Mol Biol. 2015; 111:28.6.1-28.6.11. (Methodology: Intracellular Staining/Flow Cytometry). 查看参考
      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: Intracellular Staining/Flow Cytometry). 查看参考
      7. Mai HL, Nguyen TVH, Branchereau J, et al. Interleukin-7 receptor blockade by an anti-CD127 monoclonal antibody in nonhuman primate kidney transplantation.. Am J Transplant. 2020; 20(1):101-111. (Clone-specific: Intracellular Staining/Flow Cytometry). 查看参考
      8. Mueller YM, Petrovas C, Bojczuk PM, et al. Interleukin-15 increases effector memory CD8+ t cells and NK Cells in simian immunodeficiency virus-infected macaques.. J Virol. 2005; 79(8):4877-85. (Clone-specific: Intracellular Staining/Flow Cytometry). 查看参考
      9. 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: Intracellular Staining/Flow Cytometry). 查看参考
      10. 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: Intracellular Staining/Flow Cytometry). 查看参考
      11. Rigillo A, Fuchs-Baumgartinger A, Sabattini S, et al. Ki-67 assessment-agreeability between immunohistochemistry and flow cytometry in canine lymphoma.. Vet Comp Oncol. 2021; 19(3):551-566. (Clone-specific: Immunohistochemistry, Intracellular Staining/Flow Cytometry). 查看参考
      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: Immunofluorescence, Intracellular Staining/Flow Cytometry).
      13. 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: Intracellular Staining/Flow Cytometry). 查看参考
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      571399 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.