Human beta Defensin-3 Recombinant
Categories: PDGF familyPDGF familyRecombinant Human Cytokines$70.00 – $2,700.00
Description
Accession
P81534
Source
Optimized DNA sequence encoding Humanbeta Defensin-3 mature chain was expressed in Escherichia Coli.
Molecular weight
Native humanBD-3 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately kDa. Recombinant beta Defensin-3 is a monomer protein consisting of46 amino acid residue subunits, and migrates as an approximately kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>98%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by its antimicrobial resistance toE.Coli and was determined to be <3 ug/ml.
Protein Sequence
MRIHYLLFAL LFLFLVPVPG HGGIINTLQK YYCRVRGGRC AVLSCLPKEE QIGKCSTRGR KCCRRKK
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
RecombinantHuman betaDefensin-3 was lyophilized from.2 μm filtered100mM NaCl,mM PB, pH.4.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage
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 at2° -8° 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.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Molecular function
Methods
Cryo Immunoelectron microscopy
-
Parental strains NTHI 86-028NP::
rpsL and the isogenicsapBC mutant strain were grown to mid-log phase in sBHI, normalized for cell number, and pelleted. - Pellets were resuspended in 10 mM sodium phosphate buffer pH 7.4, supplemented with 2% sBHI.
- Human LL-37 cathelicidin or human beta defensin 3 were added to cultures at a final concentration of 0.25 µg LL-37/ml or 1 µg hBD3/ml.
- Cultures containing AMPs or cells alone were incubated at 37°C, 5% CO2, static for 30 minutes.
- Samples were transfered to ice and cells were pelleted by centrifugation, washed in 100 mM PIPES pH 7.0, and then resuspended in 100 mM PIPES.
- For immunolocalization of AMPs at the ultrastructural level, bacteria were fixed in 4% paraformaldehyde/0.05% glutaraldehyde in 100 mM PIPES/0.5 mM MgCl2, pH 7.2 (or PBS) for 1 hr at 4°C.
- Samples were then embedded in 10% gelatin and infiltrated overnight with 2.3 M sucrose/20% polyvinyl…
The SapF ATPase is dispensable for resistance to the AMPs hBD-3 and LL-37.
- The parent strain and the SapF ATPase-deficient strain (A and C) or the Wild Type and SapD-ATPase deficient strain (B and D) were incubated with increasing concentrations of hBD-3 (A and B) or LL-37 (C and D) for 1 h and then plated on chocolate agar for enumeration of viable bacteria.
