Myoglobin (symbol Mb or MB or MYO) is an iron- and oxygen-binding protein found in the muscle tissue of vertebrates in general and in almost all mammals. It is related to hemoglobin, which is the iron- and oxygen-binding protein in blood, specifically in the red blood cells. Myoglobin is found in Type I muscle, Type II A and Type II B, but most texts consider myoglobin not to be found in smooth muscle. Besides, Proteasome 26S Subunit, Non ATPase 4 (PSMD4) has been identified as an interactor of MYO, thus a binding ELISA assay was conducted to detect the interaction of recombinant human MYO and recombinant human PSMD4 Briefly, MYO were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to PSMD4-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MYO 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 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of MYO and PSMD4 was shown in Figure 1, and this effect was in a dose dependent manner.
Figure. The binding activity of MYO with PSMD4.