Ki67/MKI67 Antibody (8D5) - BSA Free

Novus Biologicals | Catalog # NBP2-22112

Novus Biologicals

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rabbit

Cited:

Human, Mouse, Rabbit

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 8D5

Format

BSA Free
View all Ki67/MKI67 Primary Antibodies »

Product Specifications

Immunogen

The immunogen for this KI67/MKI67 Antibody (8D5) was made using a synthetic peptide from the internal region of Human KI67/MKI67 (amino acids: 1100-1113, 1222-1235 and 3 more spots); sequence: CEDLAGFKELFQTPG [Uniprot: P46013].

Reactivity Notes

Use in Rabbit reported in scientific literature (PMID:33691202). Human reactivity reported in the scientific literature (PMID: 23777661). Rat reactivity reported in scientific literature (PMID: 23447644). Porcine reactivity reported in scientific literature (PMID: 27046485).

Localization

Nuclear

Marker

Proliferation Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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.

Description

Novus Biologicals Knockout (KO) Validated Mouse Ki67/MKI67 Antibody (8D5) - BSA Free (NBP2-22112) is a monoclonal antibody validated for use in IHC, WB, ELISA and ICC/IF. Anti-Ki67/MKI67 Antibody: Cited in 33 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for Ki67/MKI67 Antibody (8D5) - BSA Free

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - Analysis of paraffin-embedded lung cancer (left) and rectal cancer (right) using Ki67 mouse mAb with DAB staining.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - A549 human alveolar adenocarcinoma cell line, fixation with PFA 4%, blocking (PBS, 1% BSA, 0.1% Tween), primary antibody: 1:100 in blocking buffer, O/N. Secondary antibody: AlexaFluor 594, 1 hour, RT. Counterstained with DAPI. Imaged with a fluorescence microscope. This image was submitted via customer review.
Knockout Validated: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Simple Western: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Simple Western: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - 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 317 kDa (as indicated) in the parental cell line, but is not detectable in the knockout HeLa cell line using 1:100 of Mouse Anti-Ki67/MKI67 Monoclonal Antibody (Catalog # NBP2-22112). GAPDH is shown as a loading control. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - NIH3T3 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- NBP2-22112 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - 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- NBP2-22112 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Western Blot: Ki67/MKI67 Antibody (8D5)BSA Free [NBP2-22112]

Western Blot: Ki67/MKI67 Antibody (8D5)BSA Free [NBP2-22112]

Western Blot: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - Analysis using Ki67 mouse mAb against Hela (1), MCF-7 (2) and Raji (3) cell lysate.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - U-251 MG 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-Ki67/MKI67 Antibody [8D5] conjugated to FITC (NBP2-22112F) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with anti-Ki-67 (8D5) conjugated to Alexa Fluor 488 [NBP2-22112AF488] at 10ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody (8D5) - BSA Free [NBP2-22112]

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody (8D5) [NBP2-22112] - Ki67 antibody was tested at 1:100 in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red). Image objective 40x.
Ki67/MKI67 Antibody (8D5) - BSA Free

Ki67/MKI67 (8D5) in A431 Human Cell Line -

Ki67/MKI67 (8D5) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-Ki67/MKI67 (8D5) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NBP2-22112AF488) (green) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
Ki67/MKI67 Antibody (8D5) - BSA Free

Ki67/MKI67 (8D5) in A431 Human Cell Line -

Ki67/MKI67 (8D5) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-Ki67/MKI67 (8D5) Protein-G purified Monoclonal Antibody conjugated to FITC (Catalog # NBP2-22112F) (green) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
Ki67/MKI67 Antibody (8D5) - BSA Free

Ki67/MKI67 (8D5) in A431 Human Cell Line.

Ki67/MKI67 (8D5) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-Ki67/MKI67 (8D5) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NBP2-22112AF647) (light blue) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
Ki67/MKI67 Antibody (8D5) - BSA Free

Ki67/MKI67 (8D5) in A431 Human Cell Line.

Ki67/MKI67 (8D5) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-Ki67/MKI67 (8D5) Protein-G purified Monoclonal Antibody conjugated to DyLight 488 (Catalog # NBP2-22112G) (green) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Applications for Ki67/MKI67 Antibody (8D5) - BSA Free

Application
Recommended Usage

ELISA

1:10000

Immunocytochemistry/ Immunofluorescence

1:100. Use reported by customer review

Immunohistochemistry-Frozen

reported in scientific literature (PMID 33131911)

Immunohistochemistry-Paraffin

1:200-1:1000

Western Blot

1:500-1:2000

Reviewed Applications

Read 1 review rated 5 using NBP2-22112 in the following applications:

Formulation, Preparation, and Storage

Purification

Ammonium sulfate precipitation

Formulation

PBS

Format

BSA Free

Preservative

0.03% Sodium Azide

Concentration

1.0 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

Entrez Gene IDs

4288 (Human)

Gene Symbol

MKI67

UniProt

P46013 (Human)

Additional Ki67/MKI67 Products

Product Documents for Ki67/MKI67 Antibody (8D5) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for Ki67/MKI67 Antibody (8D5) - BSA Free

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 (8D5) - BSA Free

Customer Reviews for Ki67/MKI67 Antibody (8D5) - BSA Free (1)

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Customer Images

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  • Ki-67/MKI67 Antibody (8D5)
    Name: Maxime Bourrelle-Langlois
    Application: Immunocytochemistry
    Sample Tested: A549 human alveolar adenocarcinoma cell line
    Species: Human
    Verified Customer | Posted 03/27/2018
    Ki-67/MKI67 staining of A549 human alveolar adenocarcinoma cells
    A549 human alveolar adenocarcinoma cell line, fixation with PFA 4%, blocking with PBS 1X + BSA 1% + 0.1% Tween, primary antibody used at 1:100 in blocking buffer, O/N. Secondary antibody used AlexaFluor 594, 1 hour, RT and imaged with a fluorescence microscope and counterstained with DAPI.
    Ki67/MKI67 Antibody (8D5) - BSA Free NBP2-22112

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    • A: The concentration is lot specific available upon request.

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