Ki67/MKI67 Antibody

Novus Biologicals | Catalog # NBP2-19012

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Monkey (100%), Primate (100%), Rabbit (100%), Sheep (100%). Backed by our 100% Guarantee.

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all Ki67/MKI67 Primary Antibodies »

Product Specifications

Immunogen

The immunogen for this Ki67/MKI67 Antibody was made using amino acids 1200-1250 from Human KI67/MKI67.

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: 85% in cow, guinea pig, and rhesus monkey; mole rat (80%); 75% in panda, horse, mouse, and rat; 70% homologouus in dog, and chinese hamster. Ki67/MKI67 Antibody reacted with Mouse reported in scientific literature (PMID: 27472062).

Localization

Nuclear

Marker

Proliferation Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

359 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Ki67/MKI67 Antibody

Knockout Validated: Ki67/MKI67 Antibody [NBP2-19012]

Simple Western: Ki67/MKI67 Antibody [NBP2-19012]

Simple Western: Ki67/MKI67 Antibody [NBP2-19012] - Detection of Ki67/MKI67 by Simple WesternTM. Simple Western lane view shows lysates of HeLa parental cell line and Ki67 knockout (KO) HeLa cell line. A specific band was detected for Ki67/MKI67 at approximately 312 kDa (as indicated) in the parental cell line, but is not detectable in the knockout HeLa cell line using 20 ug/mL of Rabbit Anti-Ki67/MKI67 Polyclonal Antibody (Catalog # NBP2-19012). GAPDH is shown as a loading control. This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody [NBP2-19012]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody [NBP2-19012]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody [NBP2-19012] - A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NBP2-19012 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Western Blot: Ki67/MKI67 Antibody [NBP2-19012]

Western Blot: Ki67/MKI67 Antibody [NBP2-19012]

Western Blot: Ki-67/MKI67 Antibody [NBP2-19012] - Analysis of A431 (A), HeLa (B), Ntera2 (C), and HEK293 (D) cell lysate using Ki67 antibody (NBP2-19012) at 2 ug/ml.
Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody [NBP2-19012]

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody [NBP2-19012]

Immunohistochemistry-Paraffin: Ki-67/MKI67 Antibody [NBP2-19012] - Human breast tumor stained with Ki-67 antibody (5 ug/ml), peroxidase-conjugate and DAB chromogen.
Flow Cytometry: Ki67/MKI67 Antibody [NBP2-19012]

Flow Cytometry: Ki67/MKI67 Antibody [NBP2-19012]

Flow Cytometry: Ki-67/MKI67 Antibody [NBP2-19012] - Expression in actively growing Jurkat cells: Cells were stained with 0.2 ug of Ki-67 antibody (red) and isotype control (green) and positively stained population was identified using PE conjugated goat anti-rabbit IgG secondary antibody. Cells fixed and permeabilized using ice cold 70% ethanol were used in this intracellular staining protocol.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody [NBP2-19012]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody [NBP2-19012]

Immunocytochemistry/Immunofluorescence: Ki-67/MKI67 Antibody [NBP2-19012] - Ki67 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image objective 40x. An antibody dilution of 1:10 was used.
Knockdown Validated: Ki67/MKI67 Antibody [NBP2-19012]

Knockdown Validated: Ki67/MKI67 Antibody [NBP2-19012]

Ki67-MKI67-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-19012-img0011.jpg
Ki67/MKI67 Antibody

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody [NBP2-19012] -

Downregulation of SIRT1 repressed BCa cell proliferation and induced cell cycle arrest. (a) Clone number in each well was counted and statistically analyzed in the clonogenic survival assay. ∗∗p < 0.01. (b) Clonogenic survival assay revealed cell survival of BCa cells after treatment of SIRT1-target-specific-siRNA (SIRT1 KD) and control-siRNA (NC), cultured in 6-well plates for 14 days. (c) MTT assay was used to measure the viability of BCa cells treated by SIRT1-target-specific-siRNA (SIRT1 KD, line linking squares) and negative-control-siRNA (NC, line linking circles). All shown values were mean +/- SD of three measurements and repeated three times with similar results, ∗p < 0.05. (d) Cell proliferation of BCa cells treated by SIRT1-target-specific-siRNA (B) and negative-control-siRNA (A) was assayed by Ki-67 staining (green). Nuclei (blue) were stained by DAPI. (e) Statistical analysis of percentages (%) of BCa cell populations at different stages of cell cycles. All shown values were mean +/- SD of three measurements and repeated three times with similar results. ∗p < 0.05. (f) Western blot analysis of proteins involved in G0-G1 cell cycle regulation (CDK2, CDK4, and CDK6) in the BCa cells. beta -Actin abundance was used as a control. (g) Flow cytometry analysis result for BCa cells treated with negative-control-siRNA (A) and SIRT1-target-specific-siRNA (B) for 48 h. The scale bar for (b) is 1 cm and for (d) is 40 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29147649), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Ki67/MKI67 Antibody

Application
Recommended Usage

Flow Cytometry

0.2 ug/10^6 cells

Immunocytochemistry/ Immunofluorescence

1:10

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Paraffin

5 ug/mL

Western Blot

2 ug/mL
Application Notes
Ki-67 appears to be limited to the activity phases of the cell-cycle.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS, 0.05% BSA

Preservative

0.05% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Ki67/MKI67

Ki67 is a nuclear protein associated with proliferation, named after its discovery in a Hodgkin's lymphoma-derived cell line by Scholzer and Gerdes at Kiel University, Germany (1). This proliferation marker is expressed during active phases of the cell cycle (late G1, S, G2 and M), whereas the protein remains undetectable in the G0 phase. Ki67 has two main isoforms with theoretical molecular weights of 319 kDa and 359 kDa. Its abundance is tightly regulated by synthesis and degradation with an estimated half-life of 60-90 min, regardless of its position in the cell cycle. Ki67 has been shown to be involved in ribosomal biogenesis, heterochromatin maintenance, and mitotic chromosome separation (2).

Detection of Ki67 by immunostaining is commonly used as a proliferation marker in solid tumors, as well as certain hematological malignancies (3-5). The Ki67 index, which reports on positive Ki67 stained tumor cell nuclei, has been extensively studied as a prognostic biomarker in cancers such as breast cancer and cervical cancer.

References

1. Gerdes J, Schwab U, Lemke H, Stein H. (1983) Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer. 31:13-20. PMID: 6339421

2. Starborg M, Gell K, Brundell E and Hoog C. (1996) 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. 109:143-153. 1996

3. Karamitopoulou E, Perentes E, Tolnay M, Probst A. (1998) Prognostic significance of MIB-1, p53, and bcl-2 immunoreactivity in meningiomas. Hum Pathol. 29(2):140-5. PMID: 9490273

4. Geyer FC, Rodrigues DN, Weigelt B and Reis-Filho JS. (2012) Molecular classification of estrogen receptor-positive/luminal breast cancers. Adv Anat Pathol. 19(1):39-53. PMID: 22156833

5. Ikenberg H, Bergeron C, Schmidt D, Griesser H, Alameda F, Angeloni C, Bogers J, Dachez R, Denton K, Hariri J, Keller T, von Knebel Doeberitz M, Neumann HH, Puig-Tintore LM, Sideri M, Rehm S, Ridder R; PALMS Study Group. (2013) Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study. J Natl Cancer Inst. 105(20):1550-7. PMID: 24096620

Long Name

Antigen Identified by Monoclonal Antibody Ki67

Alternate Names

Ki-67, KIA, MIB-1, MKI67, PPP1R105, TSG126, Ki67 flow cytometry, Ki-67 flow cytometry

Gene Symbol

MKI67

Additional Ki67/MKI67 Products

Product Documents for Ki67/MKI67 Antibody

Certificate of Analysis

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Product Specific Notices for Ki67/MKI67 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Cancer Biomarkers
Cell Cycle

Citations for Ki67/MKI67 Antibody

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Protocols

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FAQs for Ki67/MKI67 Antibody

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  • Q: Whats the concentration of this Ki67/MKI67 Antibody?

    A: The concentration is lot specific available upon request.

  • Q: Whats the concentration of this Ki67/MKI67 Antibody?

    A:  Optimal concentrations and conditions for each application should be determined by the user.

  • Q: Whats the concentration of this Ki67/MKI67 Antibody?

    A: The concentration is lot specific available upon request.

  • Q: Whats the concentration of this Ki67/MKI67 Antibody?

    A:  Optimal concentrations and conditions for each application should be determined by the user.

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