Optimized DNA sequence encoding Humanbeta Defensin-1mature chain was expressed in Escherichia Coli.
Native humanBD-1 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately5 kDa. Recombinant beta Defensin-1 is a monomer protein consisting of47 amino acid residue subunits, and migrates as an approximately5 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
>98%, as determined by SDS-PAGE and HPLC
The ED(50) was determined by its ability to chemoattract CD34+ dendritic cells using a concentration range of.1-1.0 ug/ml.
MRTSYLLLFT LCLLLSEMAS GGNFLTGLGH RSDHYNCVSS GGQCLYSACP IFTKIQGTCY RGKAKCCK
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
RecombinantHuman betaDefensin-1was lyophilized from.2 μm filtered100mM NaCl,mM PB, pH.4.
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Platelet and Bacterial Interaction Studies
In select studies,
- aureus was incubated in the presence of recombinant hBD-1 or hBD-1 that was captured from platelet lysates.
- Recombinant hBD-1 was also pre-incubated with an anti-hBD-1 antibody or control IgG for 1 hour prior to being added to
- Based on preliminary studies testing the effectiveness of the anti-hBD-1 antibody in blocking hBD-1 induced NET formation, a final concentration of 20 µg/ml was chosen for all studies.