Recombinant Human TGF-beta 1 GMP Protein, CF

  
  • Purity
    >97%, by SDS-PAGE with silver staining.
  • Endotoxin Level
    <0.10 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its ability to inhibit the IL-4-dependent proliferation of HT‑2 mouse T cells. Tsang, M. et al. (1995) Cytokine 7:389. The ED50 for this effect is 0.04-0.2 ng/mL.
    The specific activity of recombinant human TGF-beta 1 GMP is approximately 2.5 x 104 U/μg, which is calibrated against human TGF‑ beta 1 Standard (NIBSC code: 89/514).
  • Source
    Chinese Hamster Ovary cell line, CHO-derived Ala279-Ser390 Manufactured and tested under cGMP guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis
    Ala-Leu-Asp-Thr-Asn-Tyr-(Cys)-Phe-Ser-Ser
  • Structure / Form
    Disulfide-linked homodimer
  • Predicted Molecular Mass
    12.8 kDa (monomer)
  • SDS-PAGE
    12 kDa, reducing conditions
    24 kDa, non-reducing conditions
240-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.
Reconstitution Reconstitute at 100 μg/mL in 4 mM HCl.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
Data Images
GMP-grade Recombinant Human TGF-beta 1 (Catalog # 240-GMP) inhibits Recombinant Mouse IL‑4 (Catalog # 404-ML) induced proliferation in the HT-2 mouse T cell line. The ED50 for this effect is 0.04-0.2 ng/mL.
1 μg/lane of GMP‑grade Recombinant Human TGF-beta 1 (Catalog # 240-GMP) was resolved with SDS-PAGE under reducing (R) and nonreducing (NR) conditions and visualized by silver staining, showing single bands at 12 kDa and 24 kDa, respectively.
Background: TGF-beta 1
TGF-beta 1 (transforming growth factor beta 1) is one of three closely related mammalian members of the large TGF-beta superfamily that share a characteristic cystine knot structure (1‑7). TGF-beta 1, -2 and -3 are highly pleiotropic cytokines that are proposed to act as cellular switches that regulate processes such as immune function, proliferation and epithelial-mesenchymal transition (1‑4). Each TGF-beta isoform has some non‑redundant functions; for TGF-beta 1, mice with targeted deletion show defects in hematopoiesis and endothelial differentiation, and die of overwhelming inflammation (2). Human TGF‑ beta 1 cDNA encodes a 390 amino acid (aa) precursor that contains a 29 aa signal peptide and a 361 aa proprotein (8). A furin‑like convertase processes the proprotein to generate an N‑terminal 249 aa latency‑associated peptide (LAP) and a C‑terminal 112 aa mature TGF‑ beta 1 (8, 9). Disulfide‑linked homodimers of LAP and TGF‑ beta 1 remain non‑covalently associated after secretion, forming the small latent TGF‑ beta 1 complex (8‑10). Covalent linkage of LAP to one of three latent TGF‑ beta binding proteins (LTBPs) creates a large latent complex that may interact with the extracellular matrix (9, 10). TGF‑ beta is activated from latency by pathways that include actions of the protease plasmin, matrix metalloproteases, thrombospondin 1 and a subset of integrins (10). Mature human TGF‑ beta 1 shares 100% aa identity with pig, dog and cow TGF‑ beta 1, and 99% aa identity with mouse, rat and horse TGF‑ beta 1. It demonstrates cross‑species activity (1). TGF‑ beta 1 signaling begins with high‑affinity binding to a type II ser/thr kinase receptor termed TGF‑ beta RII. This receptor then phosphorylates and activates a second ser/thr kinase receptor, TGF‑ beta RI (also called activin receptor‑like kinase (ALK) ‑5), or alternatively, ALK‑1. This complex phosphorylates and activates Smad proteins that regulate transcription (3, 11, 12). Contributions of the accessory receptors betaglycan (also known as TGF‑ beta  RIII) and endoglin, or use of Smad‑independent signaling pathways, allow for disparate actions observed in response to TGF‑ beta in different contexts (11).
  • References:
    1. Derynck, R. and K. Miyazono (2008) Cold Spring Harbor Laboratory Press, 29.
    2. Dunker, N. and K. Krieglstein (2000) Eur. J. Biochem. 267:6982.
    3. Wahl, S.M. (2006) Immunol. Rev. 213:213.
    4. Chang, H. et al. (2002) Endocr. Rev. 23:787.
    5. Lin, J.S. et al. (2006) Reproduction 132:179.
    6. Hinck, A.P. et al. (1996) Biochemistry 35:8517.
    7. Mittl, P.R.E. et al. (1996) Protein Sci. 5:1261.
    8. Derynck, R. et al. (1985) Nature 316:701.
    9. Miyazono, K. et al. (1988) J. Biol. Chem. 263:6407.
    10. Oklu, R. and R. Hesketh (2000) Biochem. J. 352:601.
    11. de Caestecker, M. et al. (2004) Cytokine Growth Factor Rev. 15:1.
    12. Zuniga, J.E. et al. (2005) J. Mol. Biol. 354:1052.
  • Long Name:
    Transforming Growth Factor beta 1
  • Entrez Gene IDs:
    7040 (Human); 21803 (Mouse); 59086 (Rat); 397078 (Porcine); 100033900 (Equine)
  • Alternate Names:
    CEDLAP; DPD1; latency-associated peptide; TGFB; TGFB1; TGF-beta 1 protein; TGFbeta 1; TGF-beta 1; TGFbeta; TGF-beta-1; transforming growth factor beta-1; transforming growth factor, beta 1

GMP Proteins
R&D Systems' GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.

R&D Systems' quality focus includes:
  • Manufactured and tested under an ISO 9001:2008 and ISO 13485:2003 certified quality system
  • Documented processes and QA control of documentation and process changes
  • Personnel training programs
  • Raw material testing and vendor qualification/monitoring
  • Fully validated equipment, processes and test methods
  • Equipment calibration schedules using a computerized calibration program
  • Facility maintenance, safety programs and pest control
  • Material review process for variances
  • Monitoring of stability over product shelf-life

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

  • N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
  • Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial bioburden testing (using broth culture, Sabourand's dextrose and blood agar plates with results reported at 3 days and at 7 days)

Additional testing and documentation requested by the customer can be arranged at an additional cost. Testing may include, but is not limited to, USP sterility testing, positive identity testing, testing for adventitious agents and testing for residual host cell content.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.

R&D Systems sells its GMP grade recombinant protein products for research use or further manufacturing use in ex vivo cell therapy applications. They are not for in vivo use or for use as therapeutic or other drugs, biologic products or devices. Please read the following End User Terms prior to using this product.

END USER TERMS OF USE OF PRODUCT
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