Human Interferon-alpha 2a Recombinant
Categories: Interferon-IL10 familyRecombinant Human Cytokines$70.00 – $880.00
Description
Accession
P01563
Source
Optimized DNA sequence encoding Human Interferon-alpha mature chain was expressed in Escherichia Coli.
Molecular weight
Recombinant human IFN-alphaa, generated by the proteolytic removal of the signal peptide and propeptide, and has a calculated molecular mass of approximately 19 kDa. Recombinant Interferon alphaais a monomeric protein consisting of 165 amino acid residue subunits, and migrates as an approximately 19 kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity
>98%, as determined by SDS-PAGE and HPLC
Biological Activity
Theactivity was determined bya viral resistance assay of Human WISH cells, andwas found to be in the range ofx108 IU/mg.
Protein Sequence
MALTFALLVA LLVLSCKSSC SVGCDLPQTH SLGSRRTLML LAQMRKISLF SCLKDRHDFG FPQEEFGNQF QKAETIPVLH EMIQQIFNLF STKDSSAAWD ETLLDKFYTE LYQQLNDLEA CVIQGVGVTE TPLMKEDSIL AVRKYFQRIT LYLKEKKYSP CAWEVVRAEI MRSFSLSTNL QESLRSKE
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
Recombinant Interferon alphaawas lyophilized from a 0.2 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 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.
Interactor
P17181
Interactor
Biological Process
Molecular function
Methods
ATRA induces the expression and N-glycan modification of ICAM-1.
- C, U937 and WISH cells were treated with either 10 ng/ml IFN-γ or 25 µM ATRA or both.
Expression profiles of miR-29 clusters in melanoma cells.
- A375-STAT1, A375-STAT1(wt), A375 and MeWo melanoma cells were stimulated with IFN-γ for different time points.
The adipocytokines leptin, adiponectin, Nampt and NMN have no direct effects on beta-cell survival in INS-1E cells.
- INS-1E cells were exposed to cytokines (A: IL-1β, IFN-γ or TNFα) or adipocytokines (B: adiponectin, leptin, Nampt, NMN) at the indicated concentrations for 48 h and cell viability was measured by WST-1 assay.
ELISPOT assay
- An ELISPOT assay was performed to assess the IFN-γ production of autologous T cells using an IFN-γ ELISPOT kit .
- DCs were divided into four groups of 1×105 cells each, and pulsed by different ways for 12 h. Group A received GM-CSF and IL-4 only, group B received 10 μg of HSP70-HER-2-PCs purified from SKBR-3, group C received 10 μg of the HSP70-PCs purified from SKBR-3 by a method without CHAPS, and group D received 10 μg of recombinant human HSP70-HER-2 protein complex.
- After washing with PBS, the four groups of DCs were cocultured with autologous T cells isolated by a nylon wool column at a 1:10 ratio in a 96-well culture plate in the presence of 20 U/ml IL-2 for 7 days, respectively.
- The stimulated T cells (1×104/well) as effector cells and SKBR-3 cells (5×103/well) as target cells were transferred to the ELISPOT plate and incubated at 37°C…
HHV-6A Propagation and Infection
- HHV-6A strain GS 50) was determined by ocular inspection for cytopathic effect as previously described 50 was calculated according to the method of Reed and Muench −1–10−4 for 3h before they were washed with incomplete medium'>RPMI medium and further cultured in complete medium'>RPMI supplemented with IL-4 and GM-CSF in the presence of 5% CO2 and at 37°C.
- HSB-2 cells were infected in parallel as positive controls for infection.
- As positive controls for maturation, DC were cultured with 100 ng/ml lipopolysaccharide (LPS) together with 500 U/ml IFN-γ .
- To assess the effects of IFN-α, uninfected DC were stimulated with IFN-α (100–500 pg/ml, , ), and HHV-6A infected DC were cultured in the presence or absence of a neutralizing rabbit polyclonal antibody to human IFN-α (10 µg/ml; PBL interferon source, ) or normal rabbit serum ( Cruz , ,…
HHV-6A Propagation and Infection
- HHV-6A strain GS 50) was determined by ocular inspection for cytopathic effect as previously described 50 was calculated according to the method of Reed and Muench −1–10−4 for 3h before they were washed with incomplete medium'>RPMI medium and further cultured in complete medium'>RPMI supplemented with IL-4 and GM-CSF in the presence of 5% CO2 and at 37°C.
- HSB-2 cells were infected in parallel as positive controls for infection.
- As positive controls for maturation, DC were cultured with 100 ng/ml lipopolysaccharide (LPS) together with 500 U/ml IFN-γ .
- To assess the effects of IFN-α, uninfected DC were stimulated with IFN-α (100–500 pg/ml, , ), and HHV-6A infected DC were cultured in the presence or absence of a neutralizing rabbit polyclonal antibody to human IFN-α (10 µg/ml; PBL interferon source, ) or normal rabbit serum ( Cruz , ,…
Phagocytosis
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The phagocytic efficacy of macrophages isolated from healthy male and female donors was measured as in
Cn isolate) in RPMI-1640 media containing 10% human serum (50%∶50% male:female), and 100 ng/ml LPS at a density of 2×104 macrophages and incubated overnight at 37°C with 5% CO2. - All
Cn strains were grown for 2–3 d in YPD media at 37°C, washed 3× with 10 ml PBS and stained with 2 µM CMFDA for 30 minutes at 37°C. - The strains were washed 2× with 1 ml PBS, counted and opsonized with 10 µg/ml mAb 18B7 and 20% human complement for 30 minutes at 37°C before being added to the macrophages in a 1∶1 ratio in RPMI-1640 media containing 10% human serum (50%∶50% male:female), 1 µg/ml LPS and 100 ng/ml human IFN-γ .
- Macrophages and
Cn were incubated at 37°C in 5% CO2 for 2 hours. - After the incubation, the…
Not PTEN loss but IFN-γ induced PD-L1 mRNA expression in CRC cell lines.
- The relative expression level of PTEN mRNA (detected by qRT-PCR, calculated by 2−ΔΔCT method) in four groups: cells transfected with siRNA PTEN or non-specific sequences in the presence or absence of IFN-γ.
RNA Interference (RNAi)
- SiRNA transfection was carried out with lipofectamine 2000 with a concentration of 100 nM.
- Transfection efficiency was tested by qRT-PCR and western blot.
- These experiments were performed in the presence and absence of recombinant human IFN-γ (500 IU/mL, , ) and repeated in triplicates.
