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Western blot analysis of placental alkaline phosphatase (AP) was performed by loading 5ul per well of filtered cell culture supernatant obtained from cultures of Drosophila melanogaster S3 cells transfected with a Dscam-7.27.25-AP plasmid, a Dscam-1.30.30-AP plasmid, or mock transfected, and grown in 1.6ml media for 7 days, onto an 8% polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with a placental alkaline phosphatase monoclonal antibody (Product # MA5-12694) at a dilution of 1:500 overnight at 4C, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:20,000. Detection was performed using SuperSignal West Femto (Product # 35081). NOTE: The predicted molecular weights of Dscam 7.27.25-AP and Dscam 1.30.30-AP are 145.6kD and 144.4kD, respectively. Data courtesy of the Innovators Program.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2a, kappa|
|Immunogen||HEp-2 cells expressing Placental Alkaline Phosphatase (PLAP)|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||1:250-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|ELISA (ELISA)||See 1 publications below|
There are at least four distinct but related alkaline phosphatases: intestinal, placental, placental-like, and liver/bone/kidney (tissue non-specific). The first three are located together on chromosome 2 while the tissue non-specific form is located on chromosome 1. The product of this gene is a membrane bound glycosylated enzyme, also referred to as the heat stable form, that is expressed primarily in the placenta although it is closely related to the intestinal form of the enzyme as well as to the placental-like form. The coding sequence for this form of alkaline phosphatase is unique in that the 3' untranslated region contains multiple copies of an Alu family repeat. In addition, this gene is polymorphic and three common alleles (type 1, type 2 and type 3) for this form of alkaline phosphatase have been well characterized.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A vast repertoire of Dscam binding specificities arises from modular interactions of variable Ig domains.
MA5-12694 was used in ELISA to study the wide range of Dscam binding specificities arising from modular interactions of variable Ig domains
|Wojtowicz WM,Wu W,Andre I,Qian B,Baker D,Zipursky SL||Cell (130:1134)||2007|
alkaline phosphatase; Alkaline phosphatase Regan isozyme; Alkaline phosphatase, placental type; alkaline phosphomonoesterase; glycerophosphatase; Placental alkaline phosphatase 1; placental type; PLAP-1; Regan isozyme
ALP; ALPP; PALP; PLAP; PLAP-1