Detection of Mouse Alkaline Phosphatase/ALPL by Western Blot.
Western blot shows lysates of D3 mouse embryonic stem cell line, MEF mouse embryonic feeder cells, and mouse embryo tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Alkaline Phosphatase/ALPL Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2910) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for Alkaline Phosphatase/ALPL at approximately 75-80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of ALPL in Rat Stem Cells by Flow Cytometry.
Rat Mesenchymal Stem Cells were stained with Goat Anti-Mouse Alkaline Phosphatase/ALPL Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2910, filled histogram) or isotype control antibody (Catalog # AB‑108‑C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).
Alkaline Phosphatase/ALPL in Rat Mesenchymal Stem Cells.
Alkaline Phosphatase/ALPL was detected in immersion fixed rat mesenchymal stem cells using Goat Anti-Mouse Alkaline Phosphatase/ALPL Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2910) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Alkaline Phosphatase/ALPL in Mouse Embryo.
Alkaline Phosphatase/ALPL was detected in immersion fixed frozen sections of mouse embryonic (E13.5) developing vertebra using Goat Anti-Mouse Alkaline Phosphatase/ALPL Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2910) at 2 µg/mL overnight at 4 °C. Tissue was stained red and counterstained with DAPI (blue). Image courtesy of Paul J. Simmons, Ph.D., Director, Center for Stem Cell Research, The University of Texas Health Science Center at Houston, Houston, Texas, USA. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Alkaline Phosphatase/ALPL
Several distinct genes encode alkaline phosphatases (APs) in mice with different tissue-specific expression patterns. The Alpl gene, also known as Akp2, encodes the liver/bone/kidney isozyme, also known as the tissue-nonspecific AP (TNAP) (1). The Alpl gene is a key regulator of bone mineralization in mice (2). A variety of mutations in the human ALPL gene leads to different forms of hypophosphatasia, characterized by poorly mineralized cartilage and bones (3). The native ALPL is a glycosylated homodimer attached to the membrane through a GPI-anchor. The C-terminal pro peptide (residues 504 to 524) is not present in the mature form.
Terao, M. and B. Mintz (1987) Proc. Natl. Acad. Sci. USA 84:7051.
Hessle, L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:9445.
Di Mauro, S. et al. (2002) J. Bone Miner. Res. 17:1383.
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