Rat Interferon-gamma Recombinant
Category: Recombinant Rat Cytokines$70.00 – $1,000.00
Description
Accession
P01581
Source
Optimized DNA sequence encoding Rat Interferon gamma mature chain was expressed in Escherichia Coli.
Molecular weight
Native Rat gamma Interferon is generated by the proteolytic removal of the signal peptide , the molecule has a calculated molecular mass of approximately 16 kDa. Recombinant Interferon gammais a monomer protein consisting of 135 amino acid residue subunits, and migrates as an approximately 16 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the cytopathic inhibition assay with murine L929 cells infected by EMC virus, and was found to be < ng/ml, corresponding to a specific activity of >1 x units/mg.
Protein Sequence
MSATRRVLVL QLCLMALSGC YCQGTLIESL ESLKNYFNSS SMDAMEGKSL LLDIWRNWQK DGNTKILESQ IISFYLRLFE VLKDNQAISN NISVIESHLI TNFFSNSKAK KDAFMSIAKF EVNNPQIQHK AVNELIRVIH QLSPESSLRK RKRSRC
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
Recombinant Rat IFN-gamma was lyophilized from.2 μm filtered PBS solution, pH7.0.
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.
Biological Process
Biological Process
Molecular function
Methods
NO-dependent effects of leptin on PEPCK-C protein in WAT explants from rats.
- WAT explants were pre-treated or not with L-NAME (1 mmol/L) for 30 min, then exposed or not to either leptin (10 µg/L), SNAP (1 mmol/L) or INF-γ (50 µg/L) for 2 h in KRB medium containing 2% BSA.
Experiments on intraperitoneal macrophages.
- Mice at 8–12 weeks of age were injected intraperitoneally with 2 mL 3% thioglycollate 3 days before being killed by CO2 asphyxiation.
- Intraperitoneal macrophages were collected by peritoneal lavage with 10 mL sterile, cold PBS.
- Macrophages were purified by adherence to tissue culture plates for 2 h and cultured at 2 × 106 cells/mL.
- Macrophages were either incubated with 2 units/mL recombinant rat interferon-γ (IFN-γ) for 4 h followed by incubation for 24 h with or without 100 ng/mL lipopolysaccharide (LPS) in PBS vehicle, or with 400 μmol/L palmitate prepared as previously described (6/mL) were cultured with 10 ng/mL recombinant interleukin (IL)-4 ( ocky , ) for 48 h. Arginase I activity was monitored via a colorimetric assay (
Bone marrow (BM) cell cultures
- BM-derived cells were obtained by flushing tibiae and femurs.
- After lysis of red blood cells with lysing solution , the remaining cells were suspended in complete medium (CM; RPMI 1640 supplemented with 10% fetal bovine serum [FBS], antibiotics, sodium pyruvate, L-glutamine, nonessential amino acids, and 2-ME; all from, , ).
- Where indicated, the cells were stimulated with FMS-like tyrosine kinase 3 ligand (Flt3L; 200 ng/ml, , ), anti-4-1BB (5 µg/ml; clone 3H3, rat IgG2a; produced in-house), LPS (10 µg/ml), ODN-2395 (10 µg/ml), PolyI:C (10 µg/ml), TNF-α (5 ng/ml,), IFN-γ (5 ng/ml), anti-CD40 (5 µg/ml, , ), anti-μ (10 µg/ml , , ), or LA-4Ig (10 µg/ml: Bristol-Myers Squibb, , ).
HBEC support the proliferation of CD4+ and CD8+ T cells.
- For co-culture 1×105 CFSE-labelled donor PBMC were co-cultured or not with a confluent monolayer of either resting or 10 ng/ml TNF+50 ng/ml IFNγ pre-stimulated HBEC cells.
Materials
- Rat interferon-γ (IFNγ) was from.
- A methylthiazol tetrazolium (MTT) kit was from .
- Culture multi-wells and pipettes were obtained from Orange Scientific .
Materials
- Rat interferon-γ (IFNγ) was from.
- A methylthiazol tetrazolium (MTT) kit was from .
- Culture multi-wells and pipettes were obtained from Orange Scientific .
Nova1 KD increases apoptosis under basal condition and following cytokine treatment.
- Primary rat beta cells were exposed to the pro-inflammatory cytokines IL-1β + IFN-γ for 48 h and then collected for mRNA expression analyses.
