Human Alkaline Phosphatase/ALPP Antibody MAB59051: R&D Systems

Human Alkaline Phosphatase/ALPP Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Alkaline Phosphatase/ALPP in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human Alkaline Phosphatase/ALPI is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 696128
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Alkaline Phosphatase/ALPP
    Ile23-Asp506
    Accession # P05187
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Alkaline Phosphatase/ALPP by Western Blot. Western blot shows lysates of JAR human choriocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Alkaline Phosphatase/ALPP Monoclonal Antibody (Catalog # MAB59051) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Alkaline Phosphatase/ALPP at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry
Alkaline Phosphatase/ALPP in Human Placenta.

Alkaline Phosphatase/ALPP was detected in immersion fixed paraffin-embedded sections of human placenta using Mouse Anti-Human Alkaline Phosphatase/ALPP Monoclonal Antibody (Catalog # MAB59051) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to syncytiotrophoblast cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Alkaline Phosphatase/ALPP by Simple WesternTM. Simple Western lane view shows lysates of JAR human choriocarcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Alkaline Phosphatase/ALPP at approximately 74 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Alkaline Phosphatase/ALPP Monoclonal Antibody (Catalog # MAB59051). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    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/ALPP
Four distinct genes encode alkaline phosphatases (APs) in humans. The ALPP gene encodes the 67 kDa placental isozyme, also known as PLAP. In comparison, ALPI, ALPL, and ALPPL2 encode intestinal, liver/bone/kidney, and placental-like or germ cell APs, respectively. The placental alkaline phosphatase is a glycosyl phosphatidylinositol (GPI) anchored protein that is expressed on the cell surface as a homodimer. Human APs are ectopically expressed in several cancers, and their serum levels are useful tumor markers. Mature human Alkaline Phosphatase/ALPP shares 74% amino acid sequence homology with the mouse and rat enzymes.
  • Long Name:
    Alkaline Phosphatase, Placental Type
  • Entrez Gene IDs:
    250 (Human); 11650 (Mouse); 367308 (Rat)
  • Alternate Names:
    Alkaline phosphatase Regan isozyme; Alkaline Phosphatase; alkaline phosphatase, placental (Regan isozyme); alkaline phosphatase, placental type; alkaline phosphatase, placental; alkaline phosphomonoesterase; ALP; ALPP; EC 3.1.3.1; FLJ61142; glycerophosphatase; PALP; Placental alkaline phosphatase 1; PLAP; PLAP-1; Regan isozyme; SEAP
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