Human beta Tubulin I protein, His tag

Tubulin Beta (TUBb) in molecular biology can refer either to the tubulin protein superfamily of globular proteins, or one of the member proteins of that superfamily. It participate in many essential cellular processes, including mitosis. α- and β-tubulins polymerize into microtubules, a major component of the eukaryotic cytoskeleton. Both α and β tubulins have a mass of around 50kDa and are thus in a similar range compared to actin with ~42kDa. TUBb is one of six members of the tubulin superfamily, which binding drugs to kill cancerous cells by inhibiting microtubule dynamics. Besides, Myxovirus Resistance 1 (MX1) has been identified as an interactor of TUBb, thus a binding ELISA assay was conducted to detect the interaction of recombinant human TUBb and recombinant human MX1. Briefly, TUBb were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to MX1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-TUBb pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of TUBb and MX1 was in a dose dependent manner.