Detects recombinant human Alkaline Phosphatase Placental Type/ALPP and recombinant human Alkaline Phosphatase Intestinal Type/ALPI in direct ELISAs. Detects human ALPP and ALPI but not Alkaline Phosphatase Liver Type/ALPL in Western blots.
Polyclonal Sheep IgG
Mouse myeloma cell line NS0-derived recombinant human Alkaline Phosphatase/ALPP Ile23-Asp506 Accession # P05187
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.
Detection of Human Alkaline Phosphatase/ALPP/ALPI by Western Blot.
Western blot shows lysates of human placenta tissue and JEG‑3 human epithelial choriocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Alkaline Phosphatase/ALPP/ALPI Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5905) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Alkaline Phosphatase/ALPP/ALPI at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Alkaline Phosphatase/ALPP/ALPI by Simple WesternTM.
Simple Western lane view shows lysates of human placenta tissue, loaded at 0.2 mg/mL. A specific band was detected for Alkaline Phosphatase/ALPP/ALPI at approximately 76 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human Alkaline Phosphatase/ALPP/ALPI Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5905) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
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/ALPP/ALPI
ALPP (Alkaline phosphatase placental type; also SEAP, PALP-1 and ALP) is a 66-68 kDa glycoprotein member of the alkaline phosphatase family of molecules. It is principally expressed by 2nd and 3rd trimester synctiotrophoblasts and tumor cells, and represents one of four alkaline phosphatase isozymes. Experimentally, it cleaves phosphate monoesters into alcohol and phosphate. Human ALPP is synthesized as a 535 amino acid (aa) preproprecursor. It contains a 22 aa signal sequence, a 484 mature region (aa 23-506), and a 29 aa C-terminal propeptide that is cleaved to generate a GPI linkage at Asp506. The ALPP gene is highly allelic and may generate three potential splice variants that involve the signal sequence. The ALPP enzyme utilizes one Mg and two Zn ions, and functions as either a homodimer, or a heterodimer with the related Intestinal (ALPI) and Germ cell (GCAP) family members on the cell surface. ALPP is reported to oligomerize under certain conditions. Over aa 23-506, human ALPP shares 96% and 77% aa identity with chimpazeee ALPP and mouse embryonic-type alkaline phosphatase, respectively. It also shares 88% and 98% aa identity with IAP and GCAP, respectively. Over aa 23-506, human ALPP shares 86% aa identity with human ALPI.
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