Description
Human adenoviruses are widely used as models for studying cellular transformation and regulation of gene expression. The adenovirus E1A region plays a central role in each of these processes. This region of the viral genome encodes a series of related proteins with multifunctional capabilities. E1A regulates the transcriptional regulation of a wide variety of viral and cellular promoters. Most commonly, this results in the activation of transcription from the target promoter. In other cases, transcription from the targeted promoter is repressed by the action of E1A. E1A proteins also contribute to the transforming capabilities of adenoviruses. The E1A and E1B regions together comprise the transforming region of adenovirus. While expression of E1A alone is sufficient to immortalize primary cells, complete transformation requires the additional expression of the E1B region. Several other oncogenes, including the activated H-ras oncogene and polyoma middle T antigen, can complement E1A by substituting for E1B in transformation assays. Similarly, E1A can be replaced by polyoma large T antigen, c-myc and mutated p53 protein. Several conserved regions of E1A are strikingly similar to portions of other viral oncoproteins, e.g., HPV-16, HPV-18 E7 and SV40 large T antigen. Like HPV E7 and SV40 large T antigen, E1A proteins can form a specific complex with the retinoblastoma tumor suppressor gene product (Rb protein). Complex formation between the products of oncogenes and tumor suppressor genes are believed to be important in cellular transformation. The exact mechanism(s) of these complexes are not fully understood, however they may be involved in negative regulation of cellular growth and/or differentiation. M58 recognizes Adenovirus Type 5 E1A proteins.4 A trpE-E1A fusion protein was used as immunogen. The fusion protein contained the C-terminal 266 amino acids of the 13S E1A protein. This is about 90% of the E1A 13S coding sequence.
Applications include western blot analysis (1-2 µg/ml), immunoprecipitation (1-2 µg/one million cells) 4and indirect immunofluorescence of cultured cells (5-20 µg/ml). The E1A proteins migrate at various molecular weights depending on mRNA processing and posttranslational modifications; at least 60 E1A polypeptide species have been identified, ranging between 30 and 46 kDa. Two species resulting from the 12S and 13S E1A mRNAs have reduced molecular weights of 26 kDa and 32 kDa, respectively. 293 human embryonic kidney cells or E1A/Adenovirus Type 5 infected HeLa cells are suggested as positive controls.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
