| Field | Specification |
|---|---|
| Mfr No | |
| Biological Activity | |
| Endotoxin Level | |
| Formulation | |
| Molecular Weight | |
| Product Type | |
| Purity | |
| Reconstitution | |
| Source | Escherichia coli |
| Storage |
Scientific Background
Members of the Hedgehog (Hh) family are highly conserved proteins which are widely represented throughout the animal kingdom. The three known mammalian Hh proteins, Sonic (Shh), Desert (Dhh) and Indian (Ihh) are structurally related and share a high degree of amino-acid sequence identity (e.g., Shh and Ihh are 93% identical). The biologically active form of Hh molecules is obtained by autocatalytic cleavage of their precursor proteins and corresponds to approximately the N-terminal one half of the precursor molecule.
Product Description
E. coli ExpressionRecombinant Sonic HedgeHog Human protein is produced using a validated E. coli expression system and supplied as lyophilized powder for long-term stability. Suitable for use in functional bioassays, ELISA standard curves, receptor binding studies, antibody validation, and related research applications.
Protein Specifications
| Expression System | E. coli |
|---|---|
| Molecular Weight | Approximately 20.0 kDa, a single non-glycosylated polypeptide chain containing 176 amino acids. |
| Purity | >98% by SDS-PAGE and HPLC analyses. |
| Endotoxin | Less than 1EU/µg of rHuSHH as determined by LAL method. |
| Physical Form | Sterile Filtered White lyophilized (freeze-dried) powder. |
| Formulation | Lyophilized from a 0.2mm filtered concentrated solution in 20mM PB, pH 7.4, 150m… |
| Reconstitution | We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at <-20°C. Further dilutions should be made in appropriate buffered solutions. |
| Storage | This lyophilized preparation is stable at 2-8°C, but should be kept at -20°C for long term storage, … |
Functional Activity
Determined by its ability to induce alkaline phosphatase production by C3H/10T1/2 (CCL-226) cells. The expected ED50 for this effect is 0.8-1.0 μg/ml, corresponding to a Specific Activity of >1 x 103 IU/mg.
Safety & Handling
This material is offered by USA Bioworld biotech for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE. Made in China
This protein was produced in E. coli expression system. Expression system selection determines glycosylation profile, folding, and post-translational modifications. For cell-based stimulation assays, verify the expression system matches the glycosylation requirements of your target receptor or pathway.
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at <-20°C. Further dilutions should be made in appropriate buffered solutions. Reconstitute in sterile distilled water or PBS at 100 µg/mL as a standard starting concentration. Allow to dissolve at 4°C for 30 minutes without vortexing. Prepare working aliquots in 0.1% BSA carrier protein and store at −80°C.
This protein has been validated for functional bioactivity: Determined by its ability to induce alkaline phosphatase production by C3H/10T1/2 (CCL-226) cells. The expected ED50 for this effect is 0.8-1.0 μg/ml, corresponding to a Specific Activity of >1 x 103 IU/mg.. Optimal working concentrations may vary depending on your cell type, assay format, and culture conditions. Titrate the protein in a dose-response experiment to determine the optimal concentration for your system.
Endotoxin level is Less than 1EU/µg of rHuSHH as determined by LAL method. as determined by the LAL method. Low endotoxin is critical for cell-based studies because endotoxin activates NF-κB and TLR4 signalling in immune cells, producing artefactual cytokine induction that can completely mask the true biological activity of the recombinant protein.
This lyophilized preparation is stable at 2-8°C, but should be kept at -20°C for long term storage, preferably desiccated. Upon reconstitution, the preparation is stable for up to one week at 2-8°C. F Upon receipt, immediately store at −80°C. Prepare single-use working aliquots to avoid repeated freeze-thaw cycles. Lyophilized protein is stable for 6–12 months at −80°C from the date of receipt.
BioHippo offers flexible sourcing options for qualified research institutions and partners. The following may be available subject to supplier capabilities and order volume.
- Custom quantities: Bulk pricing or non-standard sizes available for high-throughput screening or scale-up projects.
- Custom formulation: Alternative reconstitution buffers or carrier proteins may be accommodated on request.
- Extended QC data: Additional bioactivity assay data, endotoxin reports, or SEC-HPLC purity profiles available on request.
Contact BioHippo customer support to discuss your requirements.