Catalogue

Background
The life cycle of a eukaryotic cell consists of various phases, two of which can morphologically and biochemically be identified. Firstly, during mitosis (M-phase), in which the cell divides into two identical daughter cells, chromosome condensation and spindle formation are microscopically visible. Secondly, in S-phase the DNA of a cell is replicated, a process that can be detected using biochemical techniques, such as the BrdU incorpoRation assay. In between the M- and S-phase two gap phases occur: the G1-phase, the gap between mitosis and the start of DNA replication, and G2-phase, the gap between completion of DNA repliCation and the onset of mitosis. From G1-phase a cell can leave the cell cycle and enter G0, a ‘quiescent’ phase. Regulation of the cell cycle predominantly occurs at three major control points, which govern the transition from G0 to G1, from G1 to S, and from G2 to M-phase.

Source
8B3G is a mouse monoclonal IgM antibody derived by fusion of SP2/0-Ag14 Mouse myeloma cells with spleen cells from a mouse immunized with a total cell lysate of the human bladder carcinoma cell line T24.

Product
Each vial contains 100 ul 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide. The product may contain some contaminating proteins next to the IgM as a result of ammonium sulphate precipitation being used for antibody purification.

Purification Method: Ammonium sulphate precipitation

Specificity
8B3G strongly stains mitotic cells and can therefore be used in flow cytometric analyses of cell suspensions to detect the mitotic index. Together with a quantitative DNA staining procedure (e.g. propidium iodide) 8B3G clearly distinguishes these M-phase cells from cell at other stages of the cell cycle (see figure). Dynamic information can be obtained by combining BrdU incorporation with 8B3G staining, which can distinguish and quantitate the four major fractions of the cell cycle.

Applications
8B3G can be used for flow cytometric analyses and immunocytochemistry. 8B3G is not suitable for immunoblotting. Optimal antibody dilution should be determined by titration; recommended range is 1:50 – 1:100 for flow cytometry, and for immunocytochemistry with avidin-biotinylated Horseradish peroxidase complex (ABC) as detection reagent.

Storage
The antibody is shipped at ambient temperature and may be stored at +4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.

Shipping Conditions: Ship at ambient temperature.

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but our company accepts no liability for any inaccuracies or omissions in this information.

Safety Datasheet(s) for this product: