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

Name: Recombinant Human TGF-beta 3 Protein, Animal-Free
Brand: R&D Systems
SKU: BT-TGFB3-AFL

R&D Systems, part of Bio-Techne | Catalog # BT-TGFB3-AFL

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Product Datasheet / COA / SDS

Key Product Details

Source:	E. coli
Accession #:	P10600.1
Applications:	Bioactivity

View Full Technical Details for Recombinant Human TGF-beta 3 Protein, Animal-Free

Technical Data
Citations / Reviews

Product Specifications for Recombinant Human TGF-beta 3 Protein, Animal-Free

Source

E. coli-derived human TGF-beta 3 protein
Ala301-Ser412
Produced using non-animal reagents in an animal-free laboratory.

Purity

>97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining

Endotoxin level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal sequence Analysis

Ala301

Predicted Molecular Mass

13 kDa

SDS-PAGE

13 kDa

Activity

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).

Scientific Data Examples for Recombinant Human TGF-beta 3 Protein, Animal-Free

	Equivalent Bioactivity of GMP,Animal-Free, and RUO gradesof Recombinant Human TGF-beta 3. 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 Protein Bioactivity. 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.
	Animal-Free™ Recombinant Human TGF‑beta 3 Protein SDS-PAGE. 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.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

BT-TGFB3-AFL

Formulation	Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in 4 mM HCl.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: TGF-beta 3

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).

Item	Value
References	Barrio, M.C. et al. (2014) Cells Tissues Organs. [Epub ahead of print; PMID 24861080]. Doetschman, T. et al. (2012) Genesis 50:59. Mittl, P.R. et al. (1996) Protein Sci. 5:1261. Sporn, M.B. (2006) Cytokine Growth Factor Rev. 17:3. Wahl, S.M. et al. (2006) Immunol. Rev. 213:213. Chang, H. et al. (2002) Endocr. Rev. 23:787. Dunker, N. and K. Krieglstein (2000) Eur. J. Biochem. 267:6982. Jin, J.Z. et al. (2014) Dev. Dyn. [Epub ahead of print; PMID 25104574]. Tang, Q.O. et al. (2009) Expert Opin. Biol Ther. 9:689. Derynck, R. et al. (1988) EMBO J. 7:3737. Miyazono, K. et al. (1988) J. Biol. Chem. 263:6407. Oklu, R. and R. Hesketh (2000) Biochem. J. 352 Pt 3:601. Cui, X.M. and C.F. Shuler (2000) Int. J. Dev. Biol. 44:397. de Caestecker, M. (2004) Cytokine Growth Factor Rev. 15:1. Nakajima, A. et al. (2007) Dev. Dyn. 236:791. Iwata, J. et al. (2012) J. Clin. Invest. 122:873. Gatza, C.E. et al. (2010) Cell. Signal. 22:1163.
Long Name	Transforming Growth Factor beta 3
Entrez Gene IDs	7043 (Human); 21809 (Mouse); 25717 (Rat)
Alternate Names	ARVD, ARVD1, FLJ16571, LDS5, RNHF, TGF-beta 3, TGF-beta-3, TGF-beta3, TGFB3, TGFbeta 3, transforming growth factor beta-3, transforming growth factor, beta 3

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

All molecular biology procedures use animal-free media and dedicated labware.
Dedicated fermentors are utilized in committed animal-free areas.

Purification

Protein purification columns are animal-free.
Bulk proteins are filtered using animal-free filters.
Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.
For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete Animal-Free Statement

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