Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Transforming Growth Factor β2 (hTGF-β2) #8406

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Source

Recombinant human TGF-β2 (hTGF-β2) Ala303-Ser414 (Accession #NP_003229) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant hTGF-β2 contains no "tags" and the nonglycosylated protein has a calculated MW of 12,719. DTT-reduced protein migrates as a 12 kDa polypeptide and the non-reduced cystine-linked homodimer migrates as a 24 kDa protein. The expected amino-terminal ALDAA of recombinant hTGF-β2 was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hTGF-β2. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hTGF-β2 was was determined by assessing inhibition of IL-4 induced HT-2 cell proliferation. The ED50 of each lot is between 0.1-0.3 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hTGF-β2 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hTGF-β2 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The inhibition of IL-4 induced proliferation in HT-2 cells treated with increasing concentrations of hTGF-β2 was assessed. After 48 hour treatment with hTGF-β2, cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT-1080 cells untreated or treated with TGF-β2 for 15 minutes, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (upper) and Smad2 (86F7) Rabbit mAb #3122 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hTGF-β2.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of 35% acetonitrile and 0.1% trifluoroacetic acid containing 20 μg BSA per 1 μg hTGF-β2. Carrier free: Lyophilized from a 0.22 μm filtered solution of 35% acetonitrile and 0.1% trifluoroacetic acid.

Background

TGF-β2 is produced by eosinophils, astrocytes, glioblastoma and other cancer derived cell types (1-6). TGF-β2 inhibits antigen-induced T-cell proliferation, inhibits epithelial cell proliferation, induces mesenchymal cell proliferation and has cell specific effects on apoptosis (1-8). TGF-β2 binds to TβRII and binding is promoted by TβRIII. TβRI then complexes with TβRII and TβRIII (3,5). Signaling involves phosphorylation of the SMAD proteins (2,3,9). TGF-β2 also activates Erk2, p38, and JNK pathways (9). Knockout of TGF-β2 in mice severely impacts heart, lung and eye development (10).

  1. Balzar, S. et al. (2005) J Allergy Clin Immunol 115, 110-7.
  2. Siegel, P.M. and Massagué, J. (2003) Nat Rev Cancer 3, 807-21.
  3. Bierie, B. and Moses, H.L. (2006) Nat Rev Cancer 6, 506-20.
  4. Hinz, S. et al. (2007) Cancer Res 67, 8344-50.
  5. Damstrup, L. et al. (1993) Br J Cancer 67, 1015-21.
  6. Constam, D.B. et al. (1992) J Immunol 148, 1404-10.
  7. Zhang, H. et al. (2008) Immunology 124, 304-14.
  8. Dufour, C. et al. (2008) Am J Physiol Endocrinol Metab 294, E794-801.
  9. Moustakas, A. and Heldin, C.H. (2009) Development 136, 3699-714.
  10. Dünker, N. and Krieglstein, K. (2000) Eur J Biochem 267, 6982-8.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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