Recombinant Human TGF-beta 3 Protein, Animal-Free

Equivalent bioactivity of GMP (BT-TGFB3-GMP), Animal-Free (Catalog # BT-TGFB3-AFL) and RUO (BT-TGFB3) grades of Recombinant Human TGF-beta 3 as measured by its ability to inhibit the IL-4-dependent proliferation ofHT‑2 mouse T cells (orange, green, red, respectively).

Animal-Free™ Recombinant Human TGF-beta 3 (Catalog # BT-TGFB3-AFL) inhibits recombinant mouse IL-4 induced proliferation in the HT-2 mouse T cell line. The ED50 for this effect is 7.50-75.0 pg/mL.

2 μg/lane of Animal-Free™ Recombinant Human TGF‑beta 3 Protein (Catalog # BT-TGFB3-AFL) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 10 kDa under reducing conditions.

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

Recombinant Human TGF-beta 3 Protein, Animal-Free 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 7.50-75.0 pg/mL.
  The specific activity of Recombinant Human TGF-beta 3 is >1.9 x 10⁷ IU/mg, which is calibrated against the Human TGF-beta 3 WHO International Standard (NIBSC code: 09/234).
• Source: E. coli-derived human TGF-beta 3 protein
  Ala301-Ser412
  Produced using non-animal reagents in an animal-free laboratory.
• Accession #: P10600.1
• N-terminal Sequence: Ala301
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Purity: >97%, by SDS-PAGE with 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: 13 kDa.
  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: 10 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA with Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining
• Reconstitution Instructions: Reconstitute at 500 μg/mL in 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 3 Protein, Animal-Free

• ARVD
• ARVD1
• FLJ16571
• LDS5
• RNHF
• TGFB3
• TGFbeta 3
• TGF-beta 3
• TGF-beta3
• TGF-beta-3
• transforming growth factor beta-3
• transforming growth factor, beta 3

Background

TGF­ beta 3 (transforming growth factor-beta 3) is a member of a TGF-beta superfamily subgroup that is defined by their structural and functional similarities (1-5). TGF-beta 3 and its closely related proteins, TGF-beta 1 and ­-beta 2, act as cellular switches to regulate immune function, cell proliferation, and epithelial­-mesenchymal transition (4, 6, 7). The non-redundant biological effects of TGF-beta 3 include involvement in palatogenesis, chondrogenesis, and pulmonary development (1, 2, 7-9). Human TGF-­ beta 3 cDNA encodes a 412 amino acid (aa) precursor that contains a 20 aa signal peptide and a 392 aa proprotein. The proprotein is processed by a furin­like convertase to generate a 220 aa latency­associated peptide (LAP) and a 112 aa mature TGF­-beta 3 (10, 11). Mature human TGF­-beta 3 shows 100%, 99%, and 98% aa identity with mouse/dog/horse, rat, and pig TGF-­ beta 3, respectively. TGF-beta 3 is secreted as a complex with LAP. This latent form of TGF-beta 3 becomes active upon cleavage by plasmin, matrix metalloproteases, thrombospondin-1, and a subset of integrins (12). TGF-beta 3 binds with high affinity to TGF-beta RII, a type II serine/threonine kinase receptor. This receptor then phosphorylates and activates type I serine/threonine kinase receptors, TGF-­ beta RI or ALK-­1, to modulate transcription through Smad phosphorylation (13-15). The divergent biological effects exerted by individual TGF-beta isoforms is dependent upon the recruitment of co-receptors (TGF-­ beta RIII and endoglin) and the subsequent initiation of Smad-­dependent or -independent signaling pathways (14, 16, 17).

1. Barrio, M.C. et al. (2014) Cells Tissues Organs. [Epub ahead of print; PMID 24861080].
2. Doetschman, T. et al. (2012) Genesis 50:59.
3. Mittl, P.R. et al. (1996) Protein Sci. 5:1261.
4. Sporn, M.B. (2006) Cytokine Growth Factor Rev. 17:3.
5. Wahl, S.M. et al. (2006) Immunol. Rev. 213:213.
6. Chang, H. et al. (2002) Endocr. Rev. 23:787.
7. Dunker, N. and K. Krieglstein (2000) Eur. J. Biochem. 267:6982.
8. Jin, J.Z. et al. (2014) Dev. Dyn. [Epub ahead of print; PMID 25104574].
9. Tang, Q.O. et al. (2009) Expert Opin. Biol Ther. 9:689.
10. Derynck, R. et al. (1988) EMBO J. 7:3737.
11. Miyazono, K. et al. (1988) J. Biol. Chem. 263:6407.
12. Oklu, R. and R. Hesketh (2000) Biochem. J. 352 Pt 3:601.
13. Cui, X.M. and C.F. Shuler (2000) Int. J. Dev. Biol. 44:397.
14. de Caestecker, M. (2004) Cytokine Growth Factor Rev. 15:1.
15. Nakajima, A. et al. (2007) Dev. Dyn. 236:791.
16. Iwata, J. et al. (2012) J. Clin. Invest. 122:873.
17. Gatza, C.E. et al. (2010) Cell. Signal. 22:1163.

Bioinformatics

• Uniprot:
  • P10600.1()