Recombinant Human TGF-beta 1 GMP Protein, CF GMP

R&D Systems, part of Bio-Techne | Catalog # 240-GMP

R&D Systems, part of Bio-Techne
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Key Product Details

Source

CHO

Accession #

P01137.2

Structure / Form

Disulfide-linked homodimer

Conjugate

Unconjugated

Applications

Bioactivity

View all TGF-beta 1 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human TGF-beta 1 protein
Ala279-Ser390
Manufactured and tested under cGMP guidelines.

Purity

>97%, by SDS-PAGE with silver staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala-Leu-Asp-Thr-Asn-Tyr-(Cys)-Phe-Ser-Ser

Predicted Molecular Mass

12.8 kDa (monomer)

SDS-PAGE

12 kDa, reducing conditions
24 kDa, non-reducing conditions

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 >1.0 x 107 units/mg, which is calibrated against the human TGF‑ beta1 Standard (NIBSC code: 89/514).

Host Cell Protein

< 0.5 ng per µg of protein when tested by ELISA.

Mycoplasma

Negative for Mycoplasma.

Scientific Data Images for Recombinant Human TGF-beta 1 GMP Protein, CF

Recombinant Human TGF-beta 1 GMP Protein Bioactivity

Recombinant Human TGF-beta 1 GMP Protein Bioactivity

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. Four independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP TGF-beta 1.
Recombinant Human TGF-beta 1 GMP Protein SDS-PAGE

Recombinant Human TGF-beta 1 GMP Protein SDS-PAGE

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.

Formulation, Preparation and Storage

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 with polar packs. 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 12 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.

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‑ beta1 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‑ beta1 (8, 9). Disulfide‑linked homodimers of LAP and TGF‑ beta1 remain non‑covalently associated after secretion, forming the small latent TGF‑ beta1 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‑ beta1 shares 100% aa identity with pig, dog and cow TGF‑ beta1, and 99% aa identity with mouse, rat and horse TGF‑ beta1. It demonstrates cross‑species activity (1). TGF‑ beta1 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

Alternate Names

TGF beta1, TGFB, TGFB1, TGFbeta 1

Entrez Gene IDs

7040 (Human); 21803 (Mouse); 59086 (Rat); 397078 (Porcine); 100033900 (Equine)

Gene Symbol

TGFB1

UniProt

P01137.2

Additional TGF-beta 1 Products

Product Documents for Recombinant Human TGF-beta 1 GMP Protein, CF

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

  • Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 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 testing according to USP
  • Host Cell Protein testing performed by ELISA
  • Mycoplasma testing by ribosomal RNA hybridization assay

Additional testing and documentation requested by the customer can be arranged at an additional cost. 

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 GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.

Product Specific Notices for Recombinant Human TGF-beta 1 GMP Protein, CF

Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

For preclinical, or clinical ex vivo use

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