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Flow cytometric analysis of Alkaline Phosphatase expression on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 33 grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested and stained with BD Horizon™ V450 Mouse anti-Human Alkaline Phosphatase antibody (solid line) or a BD Horizon™ V450 mouse IgG1, κ isotype control (Clone MOPC-21, Cat. No.560373, dashed line). Flow cytometry was performed on a BD™ LSR II flow cytometry system.


BD Horizon™ V450 Mouse anti-Human Alkaline Phosphatase

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准备和存储
商品通知
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- mTESR™1 is a trademark of StemCell Technologies.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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The B4-78 monoclonal antibody reacts with the tissue-nonspecific isozyme of alkaline phosphatase. Alkaline phosphatases are membrane-bound glycoproteins. Four isozymes of alkaline phosphatase exist in humans: placental, placental-like, intestinal, and liver/bone/kidney. Liver/bone/kidney alkaline phosphatase is also known as tissue-nonspecific alkaline phosphatase (TNAP). Human embryonic stem cells and embryonic carcinoma cells express high levels of tissue-nonspecific alkaline phosphatase that decrease upon differentiation. Genetic and biochemical studies suggest that TNAP plays a role in skeletal mineralization.
The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

研发参考 (6)
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Addison WN, Sorensen ES, Kaartinen MT, McKee MD. Pyrophosphate inhibits mineralization of osteoblast cultures by binding to mineral, up-regulating osteopontin, and inhibiting alkaline phosphatase activity. J Biol Chem. 2007; 282(21):15872-15883. (Biology). 查看参考
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Dorheim MA, Sullivan M, Dandapani V, et al. Osteoblastic gene expression during adipogenesis in hematopoietic supporting murine bone marrow stromal cells. J Cell Physiol. 1993; 154(2):317-328. (Clone-specific). 查看参考
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Eghbali-Fatourechi GZ, Lamsam J, Fraser D, Nagel D, Riggs BL, Khosla S. Circulating osteoblast-lineage cells in humans. N Engl J Med. 2005; 352(19):1959-1966. (Clone-specific). 查看参考
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International Stem Cell Initiative. Characterization of human embryonic stem cell lines by the International Stem Cell Initiative. Nat Biotechnol. 2007; 25(7):803-816. (Biology). 查看参考
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Lawson GM, Katzmann JA, Kimlinger TK, O'Brien JF. Isolation and preliminary characterization of a monoclonal antibody that interacts preferentially with the liver isoenzyme of human alkaline phosphatase. Clin Chem. 1985; 31(3):381-385. (Immunogen). 查看参考
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O'Connor MD, Kardel MD, Iosfina I, et al. Alkaline phosphatase-positive colony formation is a sensitive, specific, and quantitative indicator of undifferentiated human embryonic stem cells. Stem Cells. 2008; 26(5):1109-1116. (Biology). 查看参考
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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