Clone:
REA183
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC, IF, IHC, MC, 3D-IF
Alternative names:
MKI67, KIA, MIB-, MIB-1, PPP1R105, Ki67

Extended validation for Ki-67 Antibody, anti-human/mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA183
Ki-67++
B56++
20Raj1++
SolA15+
7B11+
Cells were incubated with an excess of purified unconjugated Ki-67 (REA183) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for Ki-67. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with a suitable counterstaining followed by a fixation and permeabilization step using the FoxP3 Staining Buffer Set. Cells were then stained with Ki-67 antibodies. As a control, Ki-67 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for Ki-67 Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA183 recognizes the Ki-67 antigen, a nuclear and nucleolar protein, which is strictly associated with cell proliferation. Two Ki-67 isoforms with molecular weight, 395 and 345 kDa are known. Both isoforms contain 16 “Ki-67 repeat“ sequences, each of which includes a conserved 66 bp “Ki-67 motif“. Expression of Ki-67 protein is present during all active phases of the cell cycle (G(1), S, G(2), and mitosis), but is absent from resting cells (G(0)). Due to strict association of Ki-67 with proliferating cells, Ki-67 is often used as an indicator of the “growth fraction“ of a given cell population. The N-terminal portion of Ki-67 contains a forkhead associated (FHA)1 domain, which is involved in interaction with proteins such as Hklp2 and NIFK.
Additional information: Clone REA183 displays negligible binding to Fc receptors.

Alternative names

MKI67, KIA, MIB-, MIB-1, PPP1R105, Ki67

Detailed product information

Technical specifications

CloneREA183
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse
AntigenKi-67
Alternative names of antigenMKI67, KIA, MIB-, MIB-1, PPP1R105, Ki67
Molecular mass of antigen [kDa]359
Entrez Gene ID4288
RRIDAB_2733585, AB_2784392, AB_2784391, AB_2784390, AB_2784389, AB_2784396, AB_2784395, AB_2752141, AB_2752089, AB_2784394, AB_2784393, AB_2784398, AB_2784397, AB_2801771, AB_2801762, AB_2652564, AB_2733584

Resources for Ki-67 Antibody, anti-human/mouse, REAfinity™

Certificates

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References for Ki-67 Antibody, anti-human/mouse, REAfinity™

Publications

  1. Verheijen, R. et al. (1989) Ki-67 detects a nuclear matrix-associated proliferation-related antigen. II. Localization in mitotic cells and association with chromosomes. J. Cell. Sci. 92: 531-540
  2. Takagi, M. et al. (2001) A novel nucleolar protein, NIFK, interacts with the forkhead associated domain of Ki-67 antigen in mitosis. J. Biol. Chem. 276: 25386-25391
  3. Bhatnagar, N. et al. (2017)
    Potential role of Vδ2
    +
    γδ T cells in regulation of immune activation in primary HIV infection.
    Front Immunol 8
  4. Campbell, S. et al. (2018) Myeloid cell recruitment versus local proliferation differentiates susceptibility from resistance to filarial infection. Elife 7(10)
  5. Schlöder, J. et al. (2017) Dimethyl fumarate therapy significantly improves the responsiveness of T cells in Multiple sclerosis patients for immunoregulation by regulatory T cells. Int J Mol Sci 18: E271
  6. Hyrenius-Wittsten, A. et al. (2018)
    De novo
    activating mutations drive clonal evolution and enhance clonal fitness in KMT2A-rearranged leukemia.
    Nat Commun. 9(1): 1770
  7. Jackson-Jones et al. (2016) Fat-associated lymphoid clusters control local IgM secretion during pleural infection and lung inflammation. Nat Commun. 7: 12651

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