Human beta Defensin-3 Recombinant

HomePDGF family, PDGF family, Recombinant Human Cytokines

Human beta Defensin-3 Recombinant

$70.00 – $2,700.00

SKU: RKP81534 Categories: PDGF family, PDGF family, Recombinant Human Cytokines Tags: bd-3, bd3, Beta-defensin 103, Beta-defensin 3, DEFB-3, DEFB103, DEFB103A, Defb3, Defensin, hBD-3, HBD3

Description

Methods (2)


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.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.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 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.
Usage	This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

Molecular function	Antimicrobial

Methods


Cryo Immunoelectron microscopy Parental strains NTHI 86-028NP::rpsL and the isogenic sapBC 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% Parental strains NTHI 86-028NP::rpsL and the isogenic sapBC 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 pyrrolidone in PIPES/MgCl₂ at 4°C. Samples were trimmed, frozen in liquid nitrogen, and sectioned with a UCT cryo-ultramicrotome . 50 nm sections were blocked with 5% FBS/5% NGS for 30 min and subsequently incubated with rabbit anti-hBD3 or rabbit anti-CAP-18 (LL-37) antibody for 1 hr at room temperature. Sections were then washed in block buffer and probed with 18 nm colloidal gold-conjugated anti-rabbit IgG (H+L) for 1 hr at room temperature. Sections were washed in PIPES buffer followed by a water rinse, and stained with 0.3% uranyl acetate/2% methyl cellulose. Samples were viewed with a JEOL 1200EX transmission electron microscope (JEOL USA Inc., Peabody, MA). All labeling experiments were conducted in parallel with controls omitting the primary antibody. These controls were consistently negative at the concentration of colloidal gold conjugated secondary antibodies used in these studies. Read more
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)… 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. Read more