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Old 04-27-2011, 06:37 PM
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Recombinant Human Follistatin 315 - 344

Background
Follistatin (FST) is a secreted glycoprotein that was first identified as a folliclestimulating hormone inhibiting substance in ovarian follicular fluid (1, 2).
Human Follistatin cDNA encodes a 344 amino acid (aa) protein with a 29 aa signal sequence, an Nterminal atypical TGF binding domain, three Follistatin domains that contain EGFlike and kazallike motifs, and a highly acidic Cterminal
tail. The first Follistatin domain (FS1) contains a heparin binding site, while FS1 and FS2 are most critical for activin binding and neutralization (3, 4). In addition to activin, Follistatin regulates bioavailability of many nonTGFβ members of the TGFβ superfamily, such as BMP6, BMP7 and myostatin (5). It also regulates hematopoietic stem cell adhesion to fibronectin via FS2, and binds angiogenin via FS2 and FS3 (6, 7).

Some Follistatin binding partners will also bind Follistatinlike proteins such as FSL3 (3, 5, 6). Of three Follistatin isoforms, the full length mature Follistatin (FST315) is the most abundant and the sole form in plasma, but has lower binding affinity for both activins and heparins than alternative isoforms (5, 8, 9). The acidic tail is missing in the splice variant FST288 which shows the highest affinity for activins, while a partial tail exists in the proteolytically produced FST303, which shows intermediate activin affinity (5, 8, 9). FST315 shares 98% aa identity with mouse, rat, equine and ovine FST, 99% with porcine and 97% with bovine FST.

Genetic deletion of Follistatin in mice, or expression of only the FST288 form, is perinatally lethal due to defects of lung, skin and musculoskeletal system
(10). Expression of only the FST315 isoform allows survival, with defects in vascularization and female fertility (10).


Description



Source
Chinese Hamster Ovary cell line, CHOderived Gly30 Trp344 Accession # P19883



Nterminal Sequence Analysis
Gly30


Predicted Molecular Mass
34.7 kDa


Specifications

SDSPAGE
40 50 kDa, reducing conditions


Activity
Measured by its ability to neutralize Activinmediated erythroid differentiation of K562 human chronic myelogenous leukemia cells. The ED50 for this effect is typically 0.01 0.05 μg/mL in the presence of 7.5 ng/mL of rhActivin A.


Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.


Purity
>95%, by SDSPAGE under reducing conditions and visualized by silver stain.


Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose and with BSA as a
carrier protein. See Certificate of Analysis for details.



Preparation and Storage


Reconstitution
Reconstitute at 100 μg/mL in sterile PBS


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 freezethaw 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.


References:
1. Shimasaki, S. et al. (1988) Proc. Natl. Acad. Sci. USA
85:4218.
2. Thompson, T.B. et al. (2005) Dev. Cell 9:535.
3. Sidis, Y. et al. (2005) Endocrinology 146:130.
4. Keutmann, H.T. et al. (2004) Mol. Endocrinol. 18:228.
5. Sidis, Y. et al. (2006) Endocrinology 147:3586.
6. MaguerSatta,
V. et al. (2006) Exp. Cell Res. 312:434.
7. Gao, X. et al. (2007) FEBS Lett. 581:5505.
8. Lerch, T.F. et al. (2007) J. Biol. Chem. 282:15930.
9. Schneyer, A.L. et al. (2004) J. Clin. Endocrinol. Metab.
89:5067.
10. Lin, SY.
et al. (2008) Mol. Endocrinol. 22:415.
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