High Mobility Group Protein 1 (HMG1) is among the most important chromatin proteins. In the nucleus HMGB1 interacts with nucleosomes, transcription factors, and histones. This nuclear protein organizes the DNA and regulates transcription. Besides, Stromal Cell Derived Factor 1 (SDF1) has been identified as an interactor of HMG1, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse HMG1 and recombinant mouse SDF1. Briefly, HMG1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to SDF1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-HMG1 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 HMG1 and SDF1 was shown in Figure 1, and this effect was in a dose dependent manner.
Figure. The binding activity of HMG1 with SDF1.