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Purified Mouse anti-Human Alkaline Phosphatase
Purified 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) grown in mTeSR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested and stained with Purified Mouse anti-Human Alkaline Phosphatase antibody (solid line) or a Purified mouse IgG1, κ isotype control (Clone MOPC-21, Cat. No. 556648, dashed line). The second step reagent was FITC goat anti-mouse Ig (Cat. No. 554001). Flow cytometry was performed on a BD LSR™ II flow cytometry system.

Purified Mouse anti-Human Alkaline Phosphatase

Immunoflourescent staining of Alkaline Phosphatase on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 52 grown in mTeSR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and stained with Purified Mouse anti-Human Alkaline Phosphatase monoclonal antibody (pseudo colored green) at 5 μg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies) and counter-staining of cell nuclei was with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.

Flow cytometric analysis of Alkaline Phosphatase expression on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) grown in mTeSR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested and stained with Purified Mouse anti-Human Alkaline Phosphatase antibody (solid line) or a Purified mouse IgG1, κ isotype control (Clone MOPC-21, Cat. No. 556648, dashed line). The second step reagent was FITC goat anti-mouse Ig (Cat. No. 554001). Flow cytometry was performed on a BD LSR™ II flow cytometry system.

Immunoflourescent staining of Alkaline Phosphatase on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 52 grown in mTeSR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and stained with Purified Mouse anti-Human Alkaline Phosphatase monoclonal antibody (pseudo colored green) at 5 μg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies) and counter-staining of cell nuclei was with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.

Product Details
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BD Pharmingen™
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), Bioimaging, Immunofluorescence (Tested During Development), Immunochemistry, Immunoprecipitation, Western blot (Reported)
0.5 mg/ml
249
AB_10643767
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. mTESR™1 is a trademark of StemCell Technologies.
561432 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.

561432 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
561432 Rev.1
Citations & References
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Development References (7)

  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: Immunocytochemistry (cytospins)). 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: Flow cytometry). 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. Karp JM, Ferreira LS, Khademhosseini A, Kwon AH, Yeh J, Langer RS. Cultivation of human embryonic stem cells without the embryoid body step enhances osteogenesis in vitro. Stem Cells. 2006; 24:835-843. (Biology). View Reference
  6. 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
  7. 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 (7) View Less
561432 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.