Recombinant Human TGF-beta 1 Avi-tag Protein, CF Summary
Additional Information |
Biotinylated |
Details of Functionality |
The biotin to protein ratio is greater than 0.7 as determined by the HABA assay. Measured by its binding ability in a functional ELISA. When Recombinant
Human TGF-beta RII Fc Chimera
(Catalog #
341-BR)
is
immobilized at 0.1 µg/mL (100 µL/well), Biotinylated Recombinant Human TGF-beta 1 Avi-tag (Catalog # AVI7754) binds with an ED 50 of 0.80-6.40
ng/mL. |
Source |
Human embryonic kidney cell, HEK293-derived human TGF-beta 1 protein Avi-tag | Human TGF-beta 1 (Ala279-Ser390) Accession # NP_000651.3 | N-terminus | C-terminus | |
|
Accession # |
|
N-terminal Sequence |
Gly of Avi-tag |
Structure / Form |
Disulfide-linked homodimer, biotinylated via Avi-tag |
Protein/Peptide Type |
Recombinant Proteins |
Purity |
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note |
<0.10 EU per 1 μg of the protein by the LAL method. |
Applications/Dilutions
Dilutions |
|
Theoretical MW |
15 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 |
12-16 kDa, under reducing conditions |
Packaging, Storage & Formulations
Storage |
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 3 months, -20 to -70 °C under sterile conditions after reconstitution.
|
Buffer |
Lyophilized from a 0.2 μm filtered solution in HCl with Trehalose. |
Purity |
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Reconstitution Instructions |
Reconstitute at 250 μg/mL in sterile 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 1 Avi-tag Protein, CF
Background
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). Our Avi-tag Biotinylated TGF-beta 1 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.
- 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.
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