Human GPC3 Protein, ECD (Extracellular Domain), Recombinant

Description

GPC3 (Glypican-3) is a member of the Glypican (GPC) family. Glypicans are a family of heparan sulfate proteoglycans that are linked to the cell surface by a glycosylphosphatidylinositol (GPI) anchor. Six members of this family have been found in mammals. All of them contain an N­terminal signal peptide, a large globular cysteine­rich domain (CRD) with 14 conserved cysteine residues, a stalk­like region containing the heparan sulfate attachment sites, and a C­terminal GPI anchoring site. GPC3 is processed by convertases that generates at least 3 ploypeptides: the 1st starts with Gln25, the 2nd starts with Ser359 after a furin cleavage site, and the 3rd starts with Val483. Therefore 2-3 different MW species are generated in the recombinant GPC3 preparation. They appear to be linked by disulfide bonds. GPC3 is involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs. GPC3 may regulate growth and tumor predisposition. Mutations in GPC3 cause a rare disorder Simpson­Golabi­Behmel Syndrome, which is characterized by pre- and post-natal overgrowth of multiple tissues and organs and an increased risk for developing embryonic tumors. GPC3 is overexpressed in various cancers including hepatocellular carcinoma and melanoma. GPC3 is implicated in regulating many signaling pathways such as IGF, FGF, BMP and Wnt. GPC3 directly binds FGF­basic by the heparan sulfate chains. GPC3 has been considered as a tumor marker and a potential target for immunotherapy. 

 

 

Gene Symbol: GPC3; GPC-3; OCI5; SGB; DGSX; MXR7; SDYS; SGBS; OCI-5; SGBS1; GTR2-2

 

NCBI Gene ID: 2719

 

Uniprot Entry: P51654

 

Construct Details: The recombinant human GPC3 extracellular or ecto-domain (ECD) is expressed as a 555 amino acid protein consisting of Gln25-Val558 region of GPC3 (Uniprot accession# P51654-3) and a C-terminal His tag, which exists as multiple proteolytically processed species under reducing conditions (see the gel image inserted).

  

Source:  Human cells stably expressing GPC3 ECD growing in chemical-defined media with no animal components or antibiotics 

 

Deduced Amino Acid Sequence:

        10         20         30         40         50         60 

QPPPPPPDAT CHQVRSFFQR LQPGLKWVPE TPVPGSDLQV CLPKGPTCCS RKMEEKYQLT  

        70         80         90        100        110        120 

ARLNMEQLLQ SASMELKFLI IQNAAVFQEA FEIVVRHAKN YTNAMFKNNY PSLTPQAFEF 

       130        140        150        160        170        180 

VGEFFTDVSL YILGSDINVD DMVNELFDSL FPVIYTQLMN PGLPDSALDI NECLRGARRD 

       190        200        210        220        230        240 

LKVFGNFPKL IMTQVSKSLQ VTRIFLQALN LGIEVINTTD HLKFSKDCGR MLTRMWYCSY  

       250        260        270        280        290        300 

CQGLMMVKPC GGYCNVVMQG CMAGVVEIDK YWREYILSLE ELVNGMYRIY DMENVLLGLF 

       310        320        330        340        350        360 

STIHDSIQYV QKNAGKLTTT IGKLCAHSQQ RQYRSAYYPE DLFIDKKVLK VAHVEHEETL 

       370        380        390        400        410        420 

SSRRRELIQK LKSFISFYSA LPGYICSHSP VAENDTLCWN GQELVERYSQ KAARNGMKNQ  

       430        440        450        460        470        480 

FNLHELKMKG PEPVVSQIID KLKHINQLLR TMSMPKGRVL DKNLDEEGFE SGDCGDDEDE 

       490        500        510        520        530        540 

CIGGSGDGMI KVKNQLRFLA ELAYDLDVDD APGNSQQATP KDNEISTFHN LGNVSTTENL 

       550 

YFQGSTGHHH HHHHH 

(Note: potential proteolytic processing Arg sites are indicated in red) 

 

M.W.: Calculated molecular mass (kDa): 61.3; Estimated by SDS-PAGE under reducing condition (kDa): 3 major species at 40, 60 and 80-100

 

Calculated PI:  6.04 

 

Calculated Extinction Coefficient (M^-1 cm^-1, at 280nm):  54290

 

Purity: >90% judged by SDS-PAGE under reducing condition (see the gel image above, labeled as DTT "+")

 

Formulation: Supplied at 0.5 mg/ml in sterile PBSpH7.4 (carrier and preservative free).

 

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

 

Biological Activity: Binds human FGF-basic/FGF2 in a functional ELISA.

 

 

 

 

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.

 

 

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 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks.

 

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

 

 

 

 

 

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

 

 

1. Nat. Genet. 12: 241 (1996)

2. J. Biol. Chem. 272:7574 (1997)

3. J. Clinical Invest. 108:497 (2001) 

4. J. Cell. Biol. 163: 625 (2003)

5. Clin Cancer Res. 10: 6612 (2004)

 

 

 

 

Product datasheet (pdf) can be downloaded here: FCL2059-PDS.pdf

 

 

 

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

 

 

 

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