Thermo Fisher Scientific Ki-67 Monoclonal Antibody (20Raj1), Brilliant Violet 650, eBioscience

Applications and Tested Dilution

• Application: Flow Cytometry (Flow)
• Tested Dilution: 5 µL (0.25 µg)/test

Product Specifications

Available Formats

• Unconjugated
• APC
• APC-eFluor 780
• Biotin
• Brilliant UV 395 / 563 / 615 / 737 / 805
• Brilliant Violet 421 / 480 / 605 / 711 / 786
• eFluor 450 / 506 / 570 / 615 / 660
• FITC
• PE / PE-Cyanine5.5 / PE-Cyanine7 / PE-eFluor 610
• PerCP-eFluor 710
• Request custom conjugation

Product Specific Information

The monoclonal antibody 20Raj1 recognizes the human Ki-67 protein, which exists in two isoforms (345 and 395 kDa) expressed in dividing cells. Ki-67 is present during active cell cycle phases (G1, S, G2, and mitosis) but absent in resting (G0) cells. During interphase, Ki-67 localizes to the nucleus, redistributing to chromosomes during mitosis, and associates with heterochromatin-bound proteins such as CBX3.

In tumor studies, Ki-67 expression is used to determine the fraction of proliferating cells. This antibody also recognizes canine Ki-67.

• Applications Reported: Intracellular staining followed by flow cytometric analysis
• Applications Tested: Pre-diluted and validated for intracellular staining of human peripheral blood cells using the Foxp3/Transcription Factor Staining Buffer Set (Product #00-5523-00)
  • Use at 5 µL (0.25 µg) per test (100 µL final volume, 10⁵–10⁸ cells/test)

Fluorochrome Information:
Brilliant Violet™ 650 (BV650) emits at 649 nm and is designed for cytometers with a 405 nm violet laser. Ensure instrument compatibility.

Buffer Recommendations:
When using multiple polymer dye-conjugated antibodies, use Super Bright Complete Staining Buffer (#SB-4401-42) or Brilliant Stain Buffer™ (#00-4409-75) to minimize non-specific interactions.

Light Sensitivity:
Protect from light to avoid photo-induced oxidation.

Fixation:
Samples can be stored in IC Fixation Buffer (#00-8222-49) or 1-step Fix/Lyse Solution (#00-5333-54) for up to 3 days at 4°C in the dark. BV650 is not compatible with methanol-based fixation.

Excitation: 407 nm
Emission: 649 nm
Laser: Violet Laser

BRILLIANT VIOLET™ is a trademark of Becton, Dickinson and Company or its affiliates, used under license. Powered by Sirigen™.

Target Information

Ki-67 is a nuclear protein expressed during late G1, S, G2, and M phases of the cell cycle. It associates with euchromatin and heterochromatin regions through its FHA domain. Expression varies topographically across the cell cycle: perinucleolar in G1, nuclear matrix in G2, and chromosomal in M phase.
Ki-67 serves as a proliferation marker, absent in G0 cells and increasing through mitosis. It is widely used to assess tumor cell proliferation and correlates strongly with tumor grade. In neuroscience, Ki-67 is also used as a marker of neuronal cell cycling and proliferation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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