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.
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
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- mTESR™1 is a trademark of StemCell Technologies.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Purified Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
556648
FITC Goat Anti-Mouse Ig RUO
Size
0.5 mg
Cat No.
554001
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Antibody Details
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
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
Development References (7)
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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). View Reference
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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: Immunocytochemistry (cytospins)). View Reference
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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: Flow cytometry). View Reference
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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). View Reference
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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
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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). View Reference
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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). View Reference
561432 Rev. 1
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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.
