Human LAP-TGFB1 Protein Complex, Biotinylated, Recombinant

Description

The TGFβ (transforming growth factor β) superfamily proteins are pleiotropic cytokines that regulate a diverse range of cellular processes, including proliferation, differentiation, migration, adhesion and apoptosis.  The TGFβ superfamily elicits 4 signaling pathways: TGFβ, Bone Morphogenic Protein (BMP), Activin, and Nodal.  Each pathway signals through a heteromeric receptor complex composed of at least 2 type-I and 2 type-II receptors with cytoplasmic serine/threonine kinase domains. The receptor complex phosphorylates and activates Smad proteins to regulate downstream gene transcription. There are 3 closely related mammalian members (TGFβ1-3) of the TGFβ family that share a characteristic cystine knot structure. They are known to function as cellular switches that regulate immune function, proliferation and epithelial­mesenchymal transition (EMT). Each TGFβ isoform has non-redundant functions. For instance, TGFβ1-deficient mice display defects in hematopoiesis and endothelial differentiation, and die of overwhelming inflammation. TGFβ1 is highly expressed in bone and acts as a stimulator of bone formation. A furin­like convertase processes it to generate an N­terminal 249 aa latency­associated peptide (LAP) and a C­terminal 112 aa mature TGFβ1. Mature human TGFβ1 is 100% identical to pig, dog and cow TGFβ1, and 99% identical to mouse, rat and horse TGFβ1, therefore showing cross­species activity. TGFβs are identified as potential therapeutic targets for cancer and cardiovascular diseases.  This gene is frequently upregulated in tumor cells, and mutations in this gene result in Camurati-Engelmann disease.

 

 

Gene Symbol: TGF-beta1; CED; LAP; DPD1; TGFB; TGFbeta

 

NCBI Gene ID: 7040

 

Uniprot Entry: P01137

 

Construct Details: The recombinant human LAP-TGFB1 complex is expressed as a 395 amino acid protein consisting of Leu30 -  Ser390 region of TGFB1 (Uniprot Accession #P01137) and a C-terminal DDK(Flag)-Myc-His tag, which exists as the disulfide-linked LAP-TGFβ1 protein (uncleaved) and non-covalently linked LAP:TGFB1 protein complex (cleaved).  

 

Source:  Human cells stably expressing LAP-TGFB1 growing in chemical-defined media with no animal components or antibiotics 

 

Deduced Amino Acid Sequence:

        10         20         30         40         50         60 

LSTCKTIDME LVKRKRIEAI RGQILSKLRL ASPPSQGEVP PGPLPEAVLA LYNSTRDRVA 

        70         80         90        100        110        120 

GESAEPEPEP EADYYAKEVT RVLMVETHNE IYDKFKQSTH SIYMFFNTSE LREAVPEPVL 

       130        140        150        160        170        180 

LSRAELRLLR LKLKVEQHVE LYQKYSNNSW RYLSNRLLAP SDSPEWLSFD VTGVVRQWLS 

       190        200        210        220        230        240 

RGGEIEGFRL SAHCSCDSRD NTLQVDINGF TTGRRGDLAT IHGMNRPFLL LMATPLERAQ 

       250        260        270        280        290        300 

HLQSSRHRRA LDTNYCFSST EKNCCVRQLY IDFRKDLGWK WIHEPKGYHA NFCLGPCPYI 

       310        320        330        340        350        360 

WSLDTQYSKV LALYNQHNPG ASAAPCCVPQ ALEPLPIVYY VGRKPKVEQL SNMIVRSCKC 

       370        380        390 

SGSTDYKDDD DKSTEQKLIS EEDLSTGHHH HHHHH 

 

 

M.W.: Calculated molecular mass 45.2 kDa; 3 major protein species by SDS-PAGE under reducing condition: ~50, ~40 and ~17 kDa corresponding to the uncleaved LAP-TGFB1(30-390aa), and cleaved LAP(30-278aa):TGFβ1(279-390aa) complex, respectively

 

Calculated PI: 7.4

 

Calculated Extinction Coefficient (M^-1 cm^-1, at 280nm): 59080

 

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 (carrier and preservative free). 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

 

Biological Activity: Binds to its receptor human ALK1 protein and anti-TGFB1 monoclonal antibodies, human IgG1 (SKU#MAB2005) and human IgG4 (SKU#MAB1561).  

 

 

 

 

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 for immediate use. 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. J. Biol. Chem. 258: 7155 (1983)

2. Nature 316: 701 (1985)

3. Annual Rev. Immunol. 16: 137 (1998)

4. Cell 75: 671 (1993).

5. Cytokine Growth Factor Rev. 18: 279 (2007).

 

 

 

 

 

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.