Ki67/MKI67 Knockout HeLa Cell Lysate

Product Discontinued

Ki67/MKI67 Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,7 bp deletion in exon 7
• Gene: MKI67

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for Ki67/MKI67 Knockout HeLa Cell Lysate

• antigen identified by monoclonal Ki-67
• Antigen Ki67
• antigen KI-67
• Ki67
• Ki-67
• KIA
• Marker Of Proliferation Ki-67
• MIB-
• MIB-1
• MKI67
• PPP1R105
• Proliferation Marker Protein Ki-67
• proliferation-related Ki-67 antigen
• Protein Phosphatase 1
• Regulatory Subunit 105
• TSG126

Background

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

FAQs for Ki67/MKI67 Lysate (NBP2-64670). (Showing 1 - of FAQ).

Bioinformatics

• Gene Symbol: MKI67