Full length (2 Exons, 86 amino acids) Recombinant HIV-1 IIIB tat produced in the E. coli Expression System. Specificity: This protein binds to murine monoclonal antibodies of defined epitope specificity and human serum polyclonal antibodies in ELISA and Western ELISA. Biological Activity: The biological specificity of this protein was determined by LTR-CAT activation in scrapie-loaded HeLe Cells, and in U373 MG cells in the presence of 100uM chlorochine diphosphate. Tat concentrations in 1ug/ml range produced at least 25 fold increase in CAT activity. This protein inhibits DP4 (CD26) activity in vitro and DP4-dependent human T-cell activation in vivo. To examine whether purified tat retained biological activity, a modified scrape loading technique was used to introduce the protein into cell monolayers. In this procedure, either a monolayer culture of HeLa cells transfected 40hr prior with plasmid pU3R-III or a monolayer culture of HeLa cell line that surface of the dish with a rubber policeman in the presence of the purified tat. Following the scraping procedure, cells were centrifuged, replated in fresh media, and CAT assays were performed 4hr after addition of protein. As graphically elevated in cells that received that tat. Applications: A dose dependent response assay should be performed to determine the optimal concentration for use in specific applications. Dilute tat stock solution in saline-citrate buffer (150mM sodium chloride, 50mM sodium citrate, pH 6.5) immediately before use. Tat readily oxidizes in buffer solutions which may change its LTR dependent transcriptional activation activity. Transcriptional activation assays with tat are performed in 1-5ug/ml range. ELISA and Western ELISA require tat in 10-100ng protein range. Storage and Stability: Aliquot to avoid repeated freezing and thawing and store at -70C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Molekulargewicht:
12
Reinheit:
Purified by ion affinity and reverse phase HPLC to 99% as determined by SDS-PAGE and HPLC. Endotoxin: 0.01EU/mg
Formulierung:
Supplied as a liquid in 50mM Tris, 0.2M KCl, pH 8.0, 5mM DTT. No stabilizers or preservatives added.
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