Alkaline Phosphatase/ALPL Antibody [Unconjugated]

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.

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.

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 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.

• Reactivity: Mu
• Applications: WB, Flow, IHC, IP, CyTOF-ready, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Alkaline Phosphatase/ALPL Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Alkaline Phosphatase/ALPL
  Phe18-Gly503
  Accession # P09242
• Specificity: Detects mouse Alkaline Phosphatase/ALPL in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 60% cross‑reactivity with recombinant human ALPL is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ALPL

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Immunoprecipitation 25 ug/mL
  • CyTOF-ready
  • Immunocytochemistry 5-15 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Alkaline Phosphatase/ALPL Antibody [Unconjugated]

• Akp2
• Alkaline phosphatase liver/bone/kidney isozyme
• alkaline phosphatase, liver/bone/kidney
• alkaline phosphatase, tissue-nonspecific isozyme
• alkaline phosphomonoesterase
• ALPL
• APTNAP
• AP-TNAP
• EC 3.1.3.1
• FLJ40094
• FLJ93059
• glycerophosphatase
• HOPS
• liver/bone/kidney-type alkaline phosphatase
• MGC161443
• tissue-nonspecific ALP
• TNAP
• TNSALP
• TNSALPMGC167935

Background

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.

1. Terao, M. and B. Mintz (1987) Proc. Natl. Acad. Sci. USA 84:7051.
2. Hessle, L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:9445.
3. Di Mauro, S. et al. (2002) J. Bone Miner. Res. 17:1383.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Alkaline Phosphatase/ALPL Antibody (AF2910)(12)

Publications using AF2910

• L Yin, ZX Zhou, M Shen, N Chen, F Jiang, SL Wang The Human Amniotic Mesenchymal Stem Cells (hAMSCs) Improve the Implant Osseointegration and Bone Regeneration in Maxillary Sinus Floor Elevation in Rabbits Stem Cells Int, 2019;2019(0):9845497. 2019 [PMID: 31885631] (ICC, Human)
• T Kazi, I Niibe, A Nishikawa, T Matsuzaki Optimal stimulation toward the dermal papilla lineage can be promoted by combined use of osteogenic and adipogenic inducers FEBS Open Bio, 2019;0(0):. 2019 [PMID: 31730301] (ICC, Mouse)
• N Charoenpha, R Aeimlapa, S Sooksawanw, J Thongbunch, J Teerapornp, S Svasti, K Wongdee Responses of primary osteoblasts and osteoclasts from hemizygous beta-globin knockout thalassemic mice with elevated plasma glucose to 1,25-dihydroxyvitamin D3 Sci Rep, 2019;9(1):13963. 2019 [PMID: 31562377] (ICC, Mouse)
• W Liu, L Zhang, K Xuan, C Hu, L Li, Y Zhang, F Jin, Y Jin Alkaline Phosphatase Controls Lineage Switching of Mesenchymal Stem Cells by Regulating the LRP6/GSK3? Complex in Hypophosphatasia Theranostics, 2018;8(20):5575-5592. 2018 [PMID: 30555565] (ICC, Human)
• MJ Carr, JS Toma, APW Johnston, PE Steadman, SA Yuzwa, N Mahmud, PW Frankland, DR Kaplan, FD Miller Mesenchymal Precursor Cells in Adult Nerves Contribute to Mammalian Tissue Repair and Regeneration Cell Stem Cell, 2018;0(0):. 2018 [PMID: 30503141] (IHC, Mouse)
• RM Sterner, KN Kremer, A Dudakovic, JJ Westendorf, AJ van Wijnen, KE Hedin Tissue-Nonspecific Alkaline Phosphatase Is Required for MC3T3 Osteoblast-Mediated Protection of Acute Myeloid Leukemia Cells from Apoptosis J. Immunol., 2018;0(0):. 2018 [PMID: 29914885] (Western Blot, Mouse)
• G Giovannell, G Giacomazzi, H Grosemans, M Sampaolesi Morphological and functional analyses of skeletal muscles from an immunodeficient animal model of limb girdle muscular dystrophy type 2E Muscle Nerve, 2018;0(0):. 2018 [PMID: 29476695] (IHC-Fr, Mouse)
• DL Coutu, KD Kokkaliari, L Kunz, T Schroeder Multicolor quantitative confocal imaging cytometry Nat. Methods, 2018;15(1):39-46. 2018 [PMID: 29320487] (IHC, Mouse)
• Arjun Deb Cardiac Fibroblasts Adopt Osteogenic Fates and Can Be Targeted to Attenuate Pathological Heart Calcification Cell Stem Cell, 2016;0(0):. 2016 [PMID: 27867037] (IHC, Mouse)
• Yoshizaki, Keigo, Hu, Lizhi, Nguyen, Thai, Sakai, Kiyoshi, He, Bing, Fong, Chak, Yamada, Yoshihik, Bikle, Daniel D, Oda, Yuko Ablation of coactivator Med1 switches the cell fate of dental epithelia to that generating hair. PLoS ONE, 2014;9(6):e99991. 2014 [PMID: 24949995] (IHC-P, Mouse)
• Isaac, J, Erthal, J, Gordon, J, Duverger, O, Sun, H-W, Lichtler, A C, Stein, G S, Lian, J B, Morasso, M I DLX3 regulates bone mass by targeting genes supporting osteoblast differentiation and mineral homeostasis in vivo. Cell Death Differ, 2014;21(9):1365-76. 2014 [PMID: 24948010] (IHC, Mouse)
• Liedert A, Mattausch L, Rontgen V, Blakytny R, Vogele D, Pahl M, Bindl R, Neunaber C, Schinke T, Harroch S, Amling M, Ignatius A Midkine-deficiency increases the anabolic response of cortical bone to mechanical loading. Bone, 2011;48(4):945-51. 2011 [PMID: 21185956] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: ALPL
• Uniprot:
  • P09242()