The secreted polypeptide noggin, encoded by the NOG gene, binds and inactivates members of the transforming growth factor-beta (TGF-beta) superfamily signaling proteins, such as bone morphogenetic protein-4 (BMP4). By diffusing through extracellular matrices more efficiently than members of the TGF-beta superfamily, noggin may have a principal role in creating morphogenic gradients. Noggin appears to have pleiotropic effect, both early in development as well as in later stages. The results of the mouse knockout of noggin suggest that it is involved in numerous developmental processes, such as neural tube fusion and joint formation. The amino acid sequence of human noggin is highly homologous to that of Xenopus, rat and mouse.
Expression System
Escherichia coli
Sequence
MQHYLHIRPAPSDNLPLVDLIEHPDPIFDPKEKDLNETLLRSLLGGHYDPGFMATSPPEDRPGGGGGAAGGAEDLAELDQLLRQRPSGAMPSEIKGLEFSEGLAQGKKQRLSKKLRRKLQMWLWSQTFCPVLYAWNDLGSRFWPRYVKVGSCFSKRSCSVPEGMVCKPSKSVHLTVLRWRCQRRGGQRCGWIPIQYPIISECKCSC with polyhistidine tag at the C-terminus.
Species
Human
Tag
polyhistidine tag at the C-terminus
Endotoxin Level
<0.1 EU per 1 μg of the protein by the LAL method.
Activity
Measure by its ability to inhibit BMP-4-induced alkaline phosphatase production by ATDC5 cells.
The ED50 for this effect is <0.05 μg/mL in the presence of 50 ng/mL of recombinant human BMP-4.
Purity
>98% as determined by SDS-PAGE. Ni-NTA chromatography
Formulation
The protein was lyophilized from a solution containing 1X PBS containing, pH 7.4.
Reconstitution
- Unopened ampoules can be stored at -20°C or -80°C.
- Need not spin the ampoule. Open the cap carefully, then dissolve the lyophilized protein with sterile water for a 100 μg/mL concentration or acc ording to the product’s Certificate of
- Analysis (CoA). (Note: Need not add carrier* to the initial reconstitution solution).
- Rinse the inner side of the ampoule gently and stand for at least 20 minutes at room temperature. (Important Note: Do Not Vortex!)
- Use the reconstituted solution immediately or store it by distributing it to aliquots and store at -20 °C. (Important Note: Avoid repeated freeze-thaw cycles)
- We recommend diluting the solution to working concentration by using buffers or medium containing a carrier*. (Important Note: Do not dilute the working solution with water, which may cause protein degradation.)
*Carriers: 0.1% BSA or 5% HSA or 10% FBS.
SDS-PAGE analysis of recombinant human Noggin