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Purified anti-ALPL Antibody

Pricing & Availability
Clone
W20113I (See other available formats)
Regulatory Status
RUO
Other Names
AP-TNAP, TNAP, Tissue-Nonspecific ALP, APTNAP, Alkaline Phosphatase, Biomineralization Associated; Alkaline Phosphatase, Tissue-Nonspecific Enzyme (TNAP); Alkaline Phosphatase, Liver/Bone/Kidney Isozyme
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
A. W20113I_PURE_ALPL_WB_1_031524
Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL of Purified anti-ALPL (clone W20113I) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • A. W20113I_PURE_ALPL_WB_1_031524
    Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL of Purified anti-ALPL (clone W20113I) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • B. W20113I_PURE_ALPL_WB_2_031524
    Saos-2 cell lysate from mock transfection (lane 1) and Saos-2 knockdown cell lysate (lane 2 and lane 3, different shRNAs transfected) were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL of Purified anti-ALPL (clone W20113I) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • C. W20113I_PURE_ALPL_ICC_040924
    Saos-2 cells and SHSY5Y cells (negative control) were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.5% Triton X-100, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 5 μg/mL of Purified anti-ALPL (clone W20113I), followed by incubation with 2.5 μg/mL of Alexa Fluor® 488 Goat anti-rat IgG (Cat. No. 405418) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). The images were captured with a 40X objective. Scale bar: 25 μm
  • D. W20113I_PURE_ALPL_IP_031524
    Whole cell extracts prepared from Saos-2 cells were immunoprecipitated for 2 hours with Purified rat IgG2b, κ isotype control (Cat. No. 400602) or 5.0 µg/test of Purified anti-ALPL (clone W20113I). The resulting IP fractions and whole cell extract input (2%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with a control antibody against a separate epitope of ALPL. Lane M: Molecular weight marker
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635951 25 µg 132€
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635952 100 µg 332€
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Description

ALPL is the tissue-nonspecific member of the alkaline phosphatase protein family, which includes four members: the tissue-nonspecific (liver/bone/kidney) ALP also called TNSALP, encoded by ALPL gene, and three tissue-specific ALPs that include intestinal (IAP), placental (PLAP-1), and placental-like 2 or germ cell (GCAP) ALPs; these enzymes are encoded by ALPI, ALPP, and ALPPL2 genes, respectively. Both ALPL and ALPI are expressed in many vertebrate species, while PLAP-1 and GCAP are expressed only in mammals. ALPL catalyzes the dephosphorylation of compounds in many transphosphorylation reactions. ALPL hydrolyzes monophosphated esters such as phosphoethanolamine, inorganic pyrophosphate, and pyridoxal 5′-phosphate.  ALPL is a homodimeric protein and each monomer possesses 524 amino acids. It is expressed mainly in liver, bone, and kidney tissues, although it can be found in all tissues. It is typically found in a membrane bound form. Its catalytic site harbors two Zn ions and one Mg ion essential for enzyme activity. Mutations in the ALPL gene causing deficiency in ALPL activity result in hypophosphatasia and skeletal defects.  ALPL regulates senescence and differentiation in mesenchymal stem cells from bone marrow. ALPL deficiency induces bone mass loss and increase in marrow fat, characteristics of premature bone ageing. ALPL was found to regulate cancer cell motility in some ovarian cancers via a non-canonical WNT pathway.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant human ALPL protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 5.0 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody (clone W20113I) does not cross-react with mouse ALPL. This product is not suggested for use in mouse.

For immunocytochemistry (ICC), we recommend using 4% PFA fixation followed by permeabilization with 0.5% Triton X-100.

This antibody (clone W20113I) is not suitable for immunohistochemistry (IHC).

Additional Product Notes

This antibody (clone W20113I) has been tested in knockout/knockdown models for WB and ICC.

RRID
AB_3106113 (BioLegend Cat. No. 635951)
AB_3106113 (BioLegend Cat. No. 635952)

Antigen Details

Structure
ALPL is a 524 amino acid protein with a predicted molecular weight of 57 kD.
Distribution

Osteoblast, bone marrow mesenchymal stem cells, liver, bone, kidney

Function
Bone and tooth mineralization, metabolism of nucleotides, sequentially hydrolyzes ATP, ADP and AMP
Interaction
Monoesters of phosphoric acid, inorganic pyrophosphate, phosphoethanolamine, and pyridoxal 5′-phosphate. Interacts with the ECM, collagen
Cell Type
Mesenchymal Stem Cells, Osteoblasts
Biology Area
Cell Biology, Stem Cells
Molecular Family
Protein Kinases/Phosphatase
Antigen References
  1. Weis MJ, et al. 1988. J Biol Chem. 263:12002-10.
  2. Kim EE and Wyckoff HW. 1991. J Mol Biol. 218:449-64.
  3. Mornet E. 2000. Human Mutation. 15:309-15.
  4. Yang Y, et al. 2012. Front Immunol. 3:314.
  5. Silvent J, et al. 2014. J Biol Chem. 289:24168-79.
  6. Liu W, et al. 2018. Bone Res. 6:27.
  7. Lou M, et al. 2019. Biochem Biophys Res Commun. 513:528-533.
Gene ID
249 View all products for this Gene ID
UniProt
View information about ALPL on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 03.15.2024

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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