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Recombinant Human Interferon-alpha 2a (IFNa2a)
(Cat. No.: C025)
Background:
At least 23 different variants of IFN-alpha are known. The individual proteins have molecular masses between 19-26 kDa and consist of proteins with lengths of 156-166 and 172 amino acids. All IFN-alpha subtypes possess a common conserved sequence region between amino acid positions 115-151 while the amino-terminal ends are variable. Many IFN-alpha subtypes differ in their sequences at only one or two positions. Naturally occurring variants also include proteins truncated by 10 amino acids at the carboxy-terminal end.
Description:
Recombinant Human IFN-alpha 2a produced in E. coli is a single, non-glycosylated, polypeptide chain containing 165 amino acids and having a molecular mass of 19241 Dalton.
Qaulity Control:
Biological activity: The specific activity as determined in a viral resistance assay using VSV-WISH cells was found to be greater than 1.0 x 108 IU/ mg.
Purity: Greater than 95% as determined by
(a) Analysis by SEC-HPLC.
(b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Molecular weight: 19.2KD+/-10% determined by reduced SDS-PAGE.
Isoelectric Point: The main zone between 5.5~6.8 analysis by IEF.
UV scan: The maximal absorption wave is 278+/-3nm.
Amino-Acid Sequence: The sequence of the first fifteen N-terminal amino acids was determined and was found to be Met-Cys-Asp-Leu-Pro-Gln-Thr-His-Ser-Leu-Gly-Ser-Arg- Arg-Thr-Leu.
Residual DNA: less than 300ng/mg analysis by solid phase blot.
Residual host cell protein: less than 0.1% analysis by ELISA.
Endotoxin: Less than 0.3ng/µg (0.3IEU/µg) determined by LAL test.
Formulation: rHuIFNa2a is lyophilized from (1mg/ml) solution containing 7.21 sodium chloride and 0.77mg ammonium acetate.
Storage: Lyophilized rHuIFN-alpha 2a although stable at room temperature for 3 weeks, should be stored desiccated below -18oC. Upon reconstitution rHuIFNα2a should be stored at 4oC between 2-7 days and for future use below -18oC. For long-term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.
Reconstitution: It is recommended to reconstitute the lyophilized rHuIFNa2a in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Latest Publications:
1: J Immunol 2009 Feb;Vol 182(3)
The Adaptor Molecule Signaling Lymphocytic Activation Molecule-Associated Protein (SAP) Regulates IFN-{gamma} and IL-4 Production in V{alpha}14 Transgenic NKT Cells via Effects on GATA-3 and T-bet Expression.
[Abstract] NKT cells comprise a rare regulatory T cell population of limited TCR diversity, with most cells using a Valpha14Jalpha18 TCR. These cells exhibit a critical dependence on the signaling adapter molecule, signaling lymphocytic activation molecule-associated protein (SAP), for their ontogeny, an aspect not seen in conventional alphabeta T cells. Prior studies demonstrate that SAP enhances TCR-induced activation of NF-kappaB in CD4(+) T cells. Because NF-kappaB is required for NKT cell development, SAP might promote the ontogeny of this lineage by signaling to NF-kappaB. In this study, we demonstrate that forced expression of the NF-kappaB target gene, Bcl-x(L), or inhibitory NF-kappaB kinase beta, a catalytic subunit of the IkappaB kinase complex essential for
NF-kappaB activation, fails to restore NKT cell development in sap(-/-) mice, suggesting that SAP mediates NKT cell development independently of NF-kappaB. To examine the role of SAP in NKT cell function, we generated NKT cells in sap(-/-) mice by expressing a transgene encoding the Valpha14Jalpha18 component of the invariant TCR. These cells bound alpha-galactosylceramide-loaded CD1d tetramers, but exhibited a very immature CD24(+)NK1.1(-) phenotype. Although sap(-/-) tetramer-reactive cells proliferated in response to TCR activation, they did not produce appreciable levels of IL-4 or IFN-gamma. The reduction in cytokine production correlated with the near absence of GATA-3 and T-bet, key transcription factors regulating cytokine expression and maturation of NKT cells. Ectopic expression of
GATA-3 partially restored IL-4 production by the NKT cells. Collectively, these data suggest that by promoting GATA-3 and T-bet expression, SAP exerts control over NKT cell development and mature NKT cell cytokine production.
2: J Immunol 2009
Feb;Vol 182(3)
MicroRNA-513 regulates B7-H1 translation and is involved in IFN-gamma-induced B7-H1 expression in cholangiocytes.
[Abstract] Biliary epithelial cells (cholangiocytes) respond to proinflammatory cytokines such as IFN-gamma and actively participate in the regulation of biliary inflammatory response in the liver. B7-H1 (also known as CD274 or PD-L1) is a member of the B7 costimulatory molecules and plays a critical immunoregulatory role in cell-mediated immune responses. In this study, we show that resting human cholangiocytes in culture express B7-H1 mRNA, but not B7-H1 protein. IFN-gamma induces B7-H1 protein expression and alters the microRNA (miRNA) expression profile in cholangiocytes. Of those IFN-gamma-down-regulated miRNAs, we identified microRNA-513 (miR-513) with complementarity to the 3'-untranslated region of B7-H1 mRNA. Targeting of the B7-H1 3'-untranslated region by
miR-513 results in translational repression. Transfection of cholangiocytes with an antisense oligonucleotide to miR-513 induces B7-H1 protein expression. Additionally, transfection of miR-513 precursor decreases IFN-gamma-induced B7-H1 protein expression and consequently influences B7-H1-associated apoptotic cell death in cocultured Jurkat cells. Thus, miR-513 regulates B7-H1 translation and is involved in IFN-gamma-induced B7-H1 expression in human cholangiocytes, suggesting a role for miRNA-mediated gene silencing in the regulation of cholangiocyte response to IFN-gamma.
3: J Immunol 2009 Feb;Vol 182(3)
Differentiated human alveolar type II cells secrete antiviral IL-29 (IFN-lambda1) in response to influenza A infection.
[Abstract] Alveolar type II epithelial cells (ATIIs) are one of the primary targets for influenza A pneumonia. The lack of a culture system for maintaining differentiated ATIIs hinders our understanding of pulmonary innate immunity during viral infection. We studied influenza A virus (IAV)-induced innate immune responses in differentiated primary human ATIIs and alveolar macrophages (AMs). Our results indicate that ATIIs, but not AMs, support productive IAV infection. Viral infection elicited strong inflammatory chemokine and cytokine responses in ATIIs, including secretion of IL-8, IL-6, MCP-1, RANTES, and MIP-1beta, but not TNF-alpha, whereas AMs secreted TNF-alpha as well as other cytokines in response to infection. Wild-type virus A/PR/8/34 induced a greater cytokine
response than reassortant PR/8 virus, A/Phil/82, despite similar levels of replication. IAV infection increased mRNA expression of IFN genes IFN-beta, IL-29 (IFN-lambda1), and IL-28A (IFN-lambda2). The major IFN protein secreted by type II cells was IL-29 and ATIIs appear to be a major resource for production of IL-29. Administration of IL-29 and IFN-beta before infection significantly reduced the release of infectious viral particles and CXC and CC chemokines. IL-29 treatment of type II cells induced mRNA expression of antiviral genes MX1, OAS, and ISG56 but not IFN-beta. IL-29 induced a dose-dependent decrease of viral nucleoprotein and an increase of antiviral genes but not IFN-beta. These results suggest that IL-29 exerts IFN-beta-independent protection in type II cells through direct
activation of antiviral genes during IAV infection.
4: J Immunol 2009 Feb;Vol 182(3)
Cutting edge: Histamine receptor H4 activation positively regulates in vivo IL-4 and IFN-gamma production by invariant NKT cells.
[Abstract] Histamine (HA) is a biogenic amine with multiple activities in the immune system. In this study we demonstrate that histamine-free histidine decarboxylase-deficient (HDC(-/-)) mice present a numerical and functional deficit in invariant NK T (iNKT) cells as evidenced by a drastic decrease of IL-4 and IFN-gamma production. This deficiency was established both by measuring cytokine levels in the serum and intracellularly among gated iNKT cells. It resulted from the lack of HA, because a single injection of this amine into HDC(-/-) mice sufficed to restore normal IL-4 and IFN-gamma production. HA-induced functional recovery was mediated mainly through the H4 histamine receptor (H4R), as assessed by its abrogation after a single injection of a selective H4R
antagonist and the demonstration of a similar iNKT cell deficit in H4R(-/-) mice. Our findings identify a novel function of HA through its H4R and suggest that it might become instrumental in modulating iNKT cell functions.
Domain Info
GeneBank Entry:
M54886
Protein Accession No.:
AAA59181
Protein Sequence:
MDLPQ THSLG SRRTL MLLAQ MRKIS LFSCL KDRHD FGFPQ EEFGN QFQKA ETIPV LHEMI QQIFN LFSTK DSSAA WDETL LDKFY TELYQ QLNDL EACVI QGVGV TETPL MKEDS ILAVR KYFQR ITLYL KEKKY SPCAW EVVRA EIMRS FSLST NLQES LRSKE
Transcript Info
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*NOTE: ALL PRODUCTS ARE FOR RESEARCH USE ONLY |
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