Recombinant Human TGF-beta 1, ACFP Protein

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human TGF-beta 1, ACFP Protein Summary

• Details of Functionality: 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 0.0400-0.200 ng/mL.
• Source: Spodoptera frugiperda, Sf 9 (baculovirus)-derived human TGF-beta 1 protein
  Ala279-Ser390
  Produced in an animal component free process (ACFP).
• Accession #: P01137
• N-terminal Sequence: Ala279
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Animal Component Free Recombinant Proteins
• Gene: TGFB1
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 12.8 kDa (monomer).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 11 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile 4 mM HCl.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TGF-beta 1, ACFP Protein

• CEDLAP
• DPD1
• latency-associated peptide
• TGF beta
• TGF beta1
• 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

Background

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).

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.

Manufacturing Process

Animal Component-Free Process (ACFP) Manufacturing Conditions
R&D Systems Animal Component-Free Process (ACFP) recombinant proteins are expressed in an animal-free certified Sf 9 insect cell line using dedicated animal-free raw materials and labware. Production and purification procedures use equipment and media that are confirmed animal-free but performed outside our dedicated animal-free laboratories. Every stage of the manufacturing process follows R&D Systems' stringent Standard Operating Procedures (SOPs). The certified Sf 9 insect cell bank has undergone extensive testing to certify the lack of cytopathogens by screening for various viruses, Mycoplasma, and Spiroplasmas using both in vitro and in vivo testing methods. For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete ACFP Statement

Bioinformatics

• Gene Symbol: TGFB1
• Uniprot:
  • P01137(Human)