Thermo Fisher Scientific Ki-67 Monoclonal Antibody (20Raj1), Brilliant Violet 605, eBioscience

Thermo Fisher Scientific Ki-67 Monoclonal Antibody (20Raj1), Brilliant Violet 605, eBioscience

Applications and Tested Dilution

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

Species Reactivity

• Dog
• Human

Antibody Information

Available Formats

• Unconjugated
• APC
• APC-eFluor 780
• Biotin
• Brilliant UV 395 / 563 / 615 / 737 / 805
• Brilliant Violet 421 / 480 / 650 / 711 / 786
• eFluor 450 / 506 / 570 / 615 / 660
• FITC
• PE / PE-Cyanine5.5 / PE-Cyanine7 / PE-eFluor 610
• PerCP-eFluor 710
• Custom conjugation available

Product Specific Information

The monoclonal antibody 20Raj1 recognizes human and canine Ki-67 protein. Two isoforms of Ki-67 (345 and 395 kDa) are expressed in dividing cells and are detectable during active phases of the cell cycle (G1, S, G2, and mitosis), but absent in resting cells (G0). During interphase, Ki-67 localizes to the nucleus and redistributes to chromosomes during mitosis.

In tumor studies, Ki-67 expression serves as a proliferation marker to determine the fraction of dividing cells.

Applications Reported:

• Intracellular staining followed by flow cytometric analysis.

Applications Tested:

• Pre-diluted and tested using intracellular staining of normal human peripheral blood cells with the Foxp3/Transcription Factor Staining Buffer Set (Product #00-5523-00).
• Recommended use: 5 µL (0.25 µg) per test (100 µL final volume, 10⁵–10⁸ cells/test).

Fluorochrome Information:

• Brilliant Violet™ 605 (BV605) emits at 605 nm, excitable with a 405 nm violet laser.
• Ensure the cytometer is compatible with this fluorochrome.

Buffer Recommendations:
When using multiple polymer dye-conjugated antibodies, use:

• Super Bright Complete Staining Buffer (Product #SB-4401-42), or
• Brilliant Stain Buffer™ (Product #00-4409-75)
  to minimize non-specific polymer interactions.

Fixation:

• Samples can be stored up to 3 days at 4°C in the dark using IC Fixation Buffer (Product #00-8222-49) or 1-step Fix/Lyse Solution (Product #00-5333-54).
• BV605 is not compatible with methanol-based fixation.

Light Sensitivity:

• Protect from light to prevent photo-induced oxidation.

Excitation/Emission:

• Excitation: 407 nm
• Emission: 605 nm
• Laser: Violet laser

Trademark Notice:
BRILLIANT VIOLET™ is a trademark of Becton, Dickinson and Company and 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 nucleolar structures through its FHA domain. Expression varies topographically through the cell cycle and is absent in G0 cells.
Ki-67 is widely used as a proliferation marker in neoplastic tissue analysis, correlating with tumor grade and growth fraction.

For Research Use Only. Not for use in diagnostic procedures.

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