Description: The monoclonal antibody 20Raj1 recognizes the human Ki-67 protein. Two isoforms of Ki-67 exist, a 345 and 395 kDa form that are expressed in dividing cells. Ki-67 is expressed in all cell types and is detectable during active phases of the cell cycle (G1, S, G2, and mitosis) but is absent from resting cells (G0). During interphase, Ki-67 expression is localized to the nucleus but redistributes to the chromosomes during mitosis and has specifically been found to associate with heterochromatin-bound proteins such as chromobox protein homolog 3 (CBX3). In studies of tumor cells, Ki-67 expression has been used as a marker for determining the fraction of proliferating cells within a given population of tumor cells.

This monoclonal antibody 20Raj1 recognizes canine Ki-67.

Applications Reported: This 20Raj1 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This 20Raj1 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins" located at Flow Protocols. This may be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Brilliant Ultra Violet™ 563 (BUV563) is a tandem dye that emits at 564 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone-specific performance should be determined empirically.

Excitation: 350 nm; Emission: 564 nm; Laser: Ultraviolet Laser.

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