Recombinant Human TGF-beta 1 GMP Protein, CF Summary
The specific activity of recombinant human TGF-beta 1 GMP is approximately 2.5 x 104 U/μg, which is calibrated against human TGF‑ beta 1 Standard (NIBSC code: 89/514).
Manufactured and tested under cGMP guidelines.
24 kDa, non-reducing conditions
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|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 at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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.
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.
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‑ 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).
- Derynck, R. and K. Miyazono (2008) Cold Spring Harbor Laboratory Press, 29.
- Dunker, N. and K. Krieglstein (2000) Eur. J. Biochem. 267:6982.
- Wahl, S.M. (2006) Immunol. Rev. 213:213.
- Chang, H. et al. (2002) Endocr. Rev. 23:787.
- Lin, J.S. et al. (2006) Reproduction 132:179.
- Hinck, A.P. et al. (1996) Biochemistry 35:8517.
- Mittl, P.R.E. et al. (1996) Protein Sci. 5:1261.
- Derynck, R. et al. (1985) Nature 316:701.
- Miyazono, K. et al. (1988) J. Biol. Chem. 263:6407.
- Oklu, R. and R. Hesketh (2000) Biochem. J. 352:601.
- de Caestecker, M. et al. (2004) Cytokine Growth Factor Rev. 15:1.
- Zuniga, J.E. et al. (2005) J. Mol. Biol. 354:1052.
Manufacturing SpecificationsGMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
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
- 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 bioburden testing (using broth culture, Sabourand's dextrose and blood agar plates with results reported at 3 days and at 7 days)
- Host Cell Protein testing performed by ELISA
- Mycoplasma testing by ribosomal RNA hybridization assay
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
The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.
TERMS AND CONDITIONS
The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.
R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.
NOTWITHSTANDING ANY OTHER PROVISIONS OF THESE TERMS AND/OR ANY OTHER AGREEMENT BETWEEN R&D SYSTEMS AND PURCHASER FOR THE PURCHASE OF THE PRODUCTS, R&D SYSTEMS' TOTAL LIABILITY TO PURCHASER ARISING FROM OR IN RELATION TO THESE TERMS, AN AGREEMENT BETWEEN THE PARTIES OR THE PRODUCTS, WHETHER ARISING IN CONTRACT, TORT OR OTHERWISE SHALL BE LIMITED TO THE TOTAL AMOUNT PAID BY PURCHASER TO R&D SYSTEMS FOR THE APPLICABLE PRODUCTS. IN NO EVENT WILL R&D SYSTEMS BE LIABLE FOR THE COST OF PROCUREMENT OF SUBSTITUTE GOODS.
Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.
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