Recombinant Cynomolgus IFNA8 Overexpression Lysate(Met1-Glu189)

• Cat.No.: IFNA8-1022CCL
• Product Overview: Recombinant Cynomolgus IFNA8 Overexpression Lysate(G7PS89) (Met1-Glu189) was expressed in HEK293 with a polyhistidine tag at the C-terminus.

Specification

• Species: Cynomolgus
• Source: HEK293
• Tag: His
• ProteinLength: Met1-Glu189
• Form: Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
• Molecular Mass: The recombinant cynomolgus IFNA8 comprises 177 amino acids and has a calculated molecular mass of 21 KDa. The apparent molecular mass of it is approximately 33 KDa respectively in SDS-PAGE.
• Storage: Store at 4°C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80°C for up to twelve months. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Case Study

Case 1: Mohammed Y, et al. Biochemistry (Mosc). 2012

Recombinant human interferon-α8 (rhIFN-α8) was produced by synthesizing an optimized gene through a two-step PCR process in Escherichia coli, addressing the high rare codon content of the human IFN-α8 gene by aligning it with E. coli's codon usage and balancing the TA-GC ratio. The successful amplification of a 504 bp DNA band corresponding to the IFN-α8 coding sequence was achieved using long overlapping primers and two Taq polymerases, with the pfu clone being essential for specific amplification. The gene was then cloned into the pBAD-TOPO vector and oriented using NcoI restriction digestion. Expression was optimized under the control of the L-arabinose-inducible P(BAD) promoter, with soluble IFN-α8 yields maximized by adjusting L-arabinose concentration, temperature, and induction time in shake flask cultures. Purification on DEAE-Sepharose yielded 100 mg/liter of IFN-α8, which was characterized by electrophoresis and immunoassays. The antiviral and anticancer activities of IFN-α8 were assessed and compared to IFN-α2a.

Fig1. SDS-PAGE analysis of rhIFN-α8 in E.coli BL21.

Fig2. MTT assay of myeloma cell line treated with different dilutions of recombinant IFN-a8 or commercial IFN-a2a compared with untreated control cells.

Case 2: García JC, et al. Biotechnol Appl Biochem. 2007

Expressing human proteins in Escherichia coli is often challenging due to codon usage differences, and typical purification methods are expensive and involve chromatography. In this study, researchers successfully expressed both human interferon alpha 2b (HuIFNα2b) and human interferon alpha 8 (HuIFNα8) in E. coli BL21-codonplus-RIL and purified them cost-effectively by passive elution from reverse-stained SDS/PAGE gels. The purified HuIFNα8 showed 1.46 times greater antiviral activity than HuIFNα2b when tested on Hep-2 human laryngeal carcinoma cells challenged with Mengo virus, demonstrating their biological activity.

Fig1. Expression of both HuIFNα subtypes in E. coli cells grown in LB medium.

Fig2. RP-HPLC analysis of purified HUIFNα8.

Application

Recombinant Cynomolgus IFNA8 protein, a member of the interferon alpha family, has a range of applications in research and potential therapeutics. This protein is known for its role in the immune response, particularly in antiviral activities. It is produced by macrophages and stimulates the production of enzymes that are key in the innate immune response to viruses and other pathogens.

In terms of applications, IFNA8 is used in various research areas, including the study of immune system responses to viral infections, the investigation of its role in cancer immunosurveillance, and the exploration of its potential as a therapeutic agent in conditions like hepatitis and certain types of cancer. The protein's ability to modulate immune responses makes it a valuable tool in understanding and potentially treating diseases where the immune system plays a critical role.

Furthermore, IFNA8 has been implicated in the regulation of glucose levels and immune reactions, suggesting its potential use in metabolic disorders and autoimmune diseases. The protein's activity in these areas is currently a subject of active research, with the goal of developing new therapeutic strategies that leverage the body's natural interferon response to combat disease.

In summary, recombinant Cynomolgus IFNA8 protein is a versatile molecule with significant applications in immunological research, antiviral therapy, and the modulation of immune responses, making it an important asset in the field of biomedicine.

Fig1. Schematic demonstrating the OCT-1 binding ability to the IFNA8 promoter region containing the rs12553612 SNP. (Hideho Okada, 2013)