Human v-Myc protein, His and GST tag

MYC (Myc proto-oncogene protein) is a nuclear phosphoprotein that binds specific sequence of DNA. MYC functions as a transcription factor and regulates transcription of target genes. It has been proven that c-Myc protein is intracellularly associated with TBP (TATA-binding protein) of the TFIID transcription initiation complex; besides, TRRAP (Transformation/transcription domain-associated protein) is thought to be an essential cofactor for the MYC. Thus a binding ELISA assay was conducted to detect the interaction of MYC with TBP and TRRAP. Briefly, recombinant human MYC were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to TBP-coated and TRRAP microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-MYC 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 MYC with TBP and TRRAP was shown in images, and this effect was in a dose dependent manner.