Human Alkaline Phosphatase/ALPP

Antibody Monoclonal Mouse IgG1 Clone # 696128 Catalog Number: MAB59051

DESCRIPTION Species Reactivity Human

Specificity Detects human Alkaline Phosphatase/ALPP in direct ELISAs. In direct ELISAs, this antibody cross­reacts at 100% with recombinant human (rh) ALPPL2, but no cross­reactivity with rhAlkaline 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 NS0­derived recombinant human Alkaline Phosphatase/ALPP Ile23­Asp506 Accession # P05187 Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (­SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS.

APPLICATIONS 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. Recommended Sample Concentration Western Blot 1 µg/mL See Below Immunohistochemistry 8­25 µg/mL See Below Simple Western 10 µg/mL See Below

DATA

Western Blot Immunohistochemistry Detection of Human Alkaline Phosphatase/ALPP by Alkaline Phosphatase/ALPP in Human Western Blot. Western blot shows lysates of JAR human Placenta. choriocarcinoma cell line. PVDF membrane was probed with Alkaline Phosphatase/ALPP was detected in 1 µg/mL of Mouse Anti­Human Alkaline Phosphatase/ALPP immersion fixed paraffin­embedded sections Monoclonal Antibody (Catalog # MAB59051) followed by HRP­ of human placenta using Mouse Anti­Human conjugated Anti­Mouse IgG Secondary Antibody (Catalog # Alkaline Phosphatase/ALPP Monoclonal HAF007). A specific band was detected for Alkaline Antibody (Catalog # MAB59051) at 15 µg/mL Phosphatase/ALPP at approximately 60 kDa (as indicated). overnight at 4 °C. Before incubation with the This experiment was conducted under reducing conditions and primary antibody, tissue was subjected to using Immunoblot Buffer Group 1. heat­induced epitope retrieval using Antigen Retrieval Reagent­Basic (Catalog # CTS013). Tissue was stained using the Anti­ Mouse HRP­DAB 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.

Simple Western 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 Anti­Human Alkaline Phosphatase/ALPP Monoclonal Antibody (Catalog # MAB59051). This experiment was conducted under reducing conditions and using the 12­230 kDa separation system. Non­specific 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 freeze­thaw cycles. l 12 months from date of receipt, ­20 to ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 6 months, ­20 to ­70 °C under sterile conditions after reconstitution.

Rev. 7/24/2018 Page 1 of 2

Human Alkaline Phosphatase/ALPP

Antibody Monoclonal Mouse IgG1 Clone # 696128 Catalog Number: MAB59051 BACKGROUND 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 placental­like 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.

Rev. 7/24/2018 Page 2 of 2