Ki-67, PE-eFluor 610, clone: 20Raj1, eBioscience™
Mouse Monoclonal Antibody
Manufacturer: Life Technologies LS61569941
Ki-67 is a nuclear protein that is expressed during various stages in the cell cycle, particularly during late G1, S, G2, and M phases. The protein has a forkhead associated domain (FHA) through which it associates with the euchromatin at the perichromosomal layer with centromeric heterochromatin, and with the nucleolus. Ki-67 is shown to have a cell cycle dependent topographical distribution, with perinucleolar expression at G1, expression in the nuclear matrix at G2, and expression on the chromosomes during M phase. Ki-67 is commonly used as a proliferation marker because it is not detected in G0 cells, but increases steadily from G1 through mitosis.Description: The monoclonal antibody 20Raj1 recognizes the human Ki-67 protein. Two isoforms of Ki-67 exist, a 345 and 395kDa 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-titrated and tested by intracellular staining of normal human peripheral blood cells. This can be used at 5 μL (0.25 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. PE-eFluor™ 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red™). Please make sure that your instrument is capable of detecting this fluorochome.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 (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) 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: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2μm post-manufacturing filtered.
|PBS with 0.2% BSA and 0.09% sodium azide; pH 7.2|
|antigen identified by monoclonal antibody Ki-67, antigen KI-67, KIA, proliferation-related Ki-67 antigen, protein phosphatase 1, regulatory subunit 105, RP11-380J17.2|
|IgG1, kappa, kappa|
|Store at 2-8°C. Do not freeze. Light-sensitive material. This tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling and experimental procedures.|
For Research Use Only.