Recombinant Human TGF-beta 3, Animal-Free Protein

TGF-beta 3 activity is determined using a TGF-beta 3-responsive firefly luciferase reporter in HEK293T cells. Cells are treated in triplicate with a serial dilution of TGF-beta 3 for 6 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. EC50 = 50 pg/ml (1.97 pM).

TGF beta 3 migrates as a single band at 25 kDa in non-reducing (NR) conditions and 13 kDa upon reduction (R). No contaminating protein bands are visible.Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+ beta -mercaptoethanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250.

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

Recombinant Human TGF-beta 3, Animal-Free Protein Summary

• Details of Functionality: No significant difference between EC₅₀ of reference and test lots
• Source: E. coli-derived human TGF-beta 3 protein
• Accession #: P10600.1
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 12.7 kDa (monomer), 25.4 kDa (dimer).
  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: Dimeric TGF- beta 3 protein only

Packaging, Storage & Formulations

• Storage: Store lyophilized protein between -20 and -80 °C until the date of expiry. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from acetonitrile/TFA
• Reconstitution Instructions: Resuspend in 10 mM HCl at >100 µg/mL, prepare single use aliquots, add carrier protein if desired.

Notes

The above product was manufactured, tested and released by R&D System's contract manufacturer, Qkine Ltd, at 1 Murdoch House, Cambridge, UK, CB5 8HW. The product is for research use only and not for the diagnostic or theraputic use.

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

Alternate Names for Recombinant Human TGF-beta 3, Animal-Free Protein

• 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 furin-­like convertase to generate a 220 aa latency-­associated 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 latent complex. This latent form of TGF-beta 3 is activated by integrins, thrombospondin-1, plasmin, and matrix metalloproteases (12, 13). It can also be activated by extreme pH and reactive oxygen species (1-5, 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 (14-16). 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 (15, 17, 18).

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. Munger, J.S. et al. (1997) Kidney Int 51:1376.
13. Wipff, P.J. and B. Hinz (2008) Eur J Cell Biol 87:601.
14. Cui, X.M. and C.F. Shuler (2000) Int. J. Dev. Biol. 44:397.
15. de Caestecker, M. (2004) Cytokine Growth Factor Rev. 15:1.
16. Nakajima, A. et al. (2007) Dev. Dyn. 236:791.
17. Iwata, J. et al. (2012) J. Clin. Invest. 122:873.
18. Gatza, C.E. et al. (2010) Cell. Signal. 22:1163.

Bioinformatics

• Uniprot:
  • P10600.1()