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V450 Mouse anti-Human Alkaline Phosphatase
V450 Mouse anti-Human Alkaline Phosphatase

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.

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.

Product Details
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BD Horizon™
Bone/Kidney/Liver Alkaline Phosphatase, TNAP, TNSALP, AP-TNAP, ALPL
Human (QC Testing), Mouse (Lack of Reactivity Confirmed in Development)
Mouse BALB/c IgG1, κ
Human Bone Alkaline Phosphatase
Flow cytometry (Routinely Tested)
5 µl
249
AB_10926207
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.

Product Notices

  1. 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).
  2. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. mTESR™1 is a trademark of StemCell Technologies.
  6. 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.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561502 Rev. 1
Antibody Details
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B4-78

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.

561502 Rev. 1
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
V450
Violet 405 nm
405 nm
450 nm
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Citations & References
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Development References (6)

  1. 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). View Reference
  2. 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). View Reference
  3. 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). View Reference
  4. 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). View Reference
  5. 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). View Reference
  6. 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). View Reference
View All (6) View Less
561502 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.