Active Syntaxin 1A, Brain (STX1A)

Syntaxin 1A (STX1A), a member of the syntaxin family ,is a nervous system-specific membrane protein responsible for fusion.Synaptosomal Associated Protein 25kDa (SNAP25) binds to STX1A to form the SNARE complex , which is essential for the release of neurotransmitters.Thus a functional ELISA assay was conducted to detect the interaction of recombinant human STX1A and recombinant human SNAP25. Briefly, STX1A was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to SNAP25-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-STX1A pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant human STX1A and recombinant human SNAP25 was shown in Figure 1, the EC50 for this effect is 0.026ug/mL.