Human ALPP/PLAP/SEAP Protein, ECD (Extracellular Domain), Fc-fusion, Biotinylated, Recombinant

Description

SEAP (secreted embryonic alkaline phosphatase) is a reporter widely used to study promoter activity or gene expression. SEAP is a truncated form of human placental alkaline phosphatase (PLAP/ALPP) by deletion of the GPI anchor. PLAP/ALPP is a metalloenzyme that catalyzes the hydrolysis of phosphoric acid monoesters. It belongs to a multigene family composed of four alkaline phosphatase isoenzymes. The enzyme functions as a homodimer and has a catalytic site containing one magnesium and two zinc ions, which are required for its enzymatic function. The protein is primarily expressed in placental and endometrial tissue.  Strong ectopic expression has been detected in ovarian adenocarcinoma, serous cystadenocarcinoma, and other ovarian cancer cells. PLAP is extremely heat stable and resistant to the inhibitor such as L-homoarginine. SEAP is secreted into cell culture supernatant and offers many advantages over intracellular reporters.  It reduces background alkaline phosphatase activity and facilitates the automation of the sampling and assay processes. It does not require the preparation of cell lysates and thus allows the continuous determination of reporter activity for kinetic analysis without disturbing the cells. There are many commercially available systems that may be employed to assay SEAP activity. 

 

 

 

Gene Symbol: ALPP; PLAP; SEAP; ALP; PALP; PLAP-1

 

NCBI Gene ID: 250

 

Uniprot Entry: P05187 

 

Construct Details: The recombinant human ALPP-Fc fusion protein is expressed as a 705 amino acid protein consisting of Ilu23 - Ala499 region of ALPP and a C-terminal Fc fusion from human IgG1, which exists as a dimer under non-reducing condition.

   

Source: Human cells stably expressing ALPP-Fc and growing in chemical-defined media with no animal component or antibiotics

 

Deduced Amino Acid Sequence: 

        10         20         30         40         50         60 

IIPVEEENPD FWNREAAEAL GAAKKLQPAQ TAAKNLIIFL GDGMGVSTVT AARILKGQKK 

        70         80         90        100        110        120 

DKLGPEIPLA MDRFPYVALS KTYNVDKHVP DSGATATAYL CGVKGNFQTI GLSAAARFNQ  

       130        140        150        160        170        180 

CNTTRGNEVI SVMNRAKKAG KSVGVVTTTR VQHASPAGTY AHTVNRNWYS DADVPASARQ 

       190        200        210        220        230        240 

EGCQDIATQL ISNMDIDVIL GGGRKYMFRM GTPDPEYPDD YSQGGTRLDG KNLVQEWLAK 

       250        260        270        280        290        300 

RQGARYVWNR TELMQASLDP SVTHLMGLFE PGDMKYEIHR DSTLDPSLME MTEAALRLLS 

       310        320        330        340        350        360 

RNPRGFFLFV EGGRIDHGHH ESRAYRALTE TIMFDDAIER AGQLTSEEDT LSLVTADHSH 

       370        380        390        400        410        420 

VFSFGGYPLR GSSIFGLAPG KARDRKAYTV LLYGNGPGYV LKDGARPDVT ESESGSPEYR 

       430        440        450        460        470        480 

QQSAVPLDEE THAGEDVAVF ARGPQAHLVH GVQEQTFIAH VMAFAACLEP YTACDLASTG 

       490        500        510        520        530        540 

THTCPPCPAP ELLGGPSVFL FPPKPKDTLM ISRTPEVTCV VVDVSHEDPE VKFNWYVDGV 

       550        560        570        580        590        600 

EVHNAKTKPR EEQYNSTYRV VSVLTVLHQD WLNGKEYKCK VSNKALPAPI EKTISKAKGQ  

       610        620        630        640        650        660 

PREPQVYTLP PSREEMTKNQ VSLTCLVKGF YPSDIAVEWE SNGQPENNYK TTPPVLDSDG 

       670        680        690        700 

SFFLYSKLTV DKSRWQQGNV FSCSVMHEAL HNHYTQKSLS LSPGK 

 

   

M.W.: Calculated molecular mass 77.7 kDa; estimated by SDS-PAGE under reducing condition: ~80-85 kDa 

 

Calculated PI: 6.12

 

Calculated extinction coefficients (M-1 cm-1, at 280nm):  83365

  

Purity: >95% judged by SDS-PAGE under reducing condition (see the gel image above, labeled as "S")

 

Formulation: Supplied at 0.5 mg/ml in sterile PBS pH7.4 (concentration determined by UV spectrometry and verified by SDS-PAGE and Coomassie blue staining). The purified recombinant protein was labeled with Biotin (3-5 Biotin per molecule) using the standard procedure.

 

Endotoxin Level: <0.1 EU per 1 μg of purified recombinant protein determined by the LAL method

 

 

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 6 months. 

 

 

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

 

 

 

Additional supporting documents, including PDS, COA and MSDS are available upon request.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.