Active Vesicle Associated Membrane Protein 2 (VAMP2)

VAMP2, also known as synaptobrevin-2, is a protein that in humans is a member of the vesicle-associated membrane protein (VAMP) family, which is part of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex. VAMP2 is involved in the regulation of neurotransmitter release from synaptic vesicles by mediating vesicle fusion with the presynaptic membrane. It is essential for synaptic vesicle exocytosis and neurotransmitter release, playing a key role in synaptic transmission. Syntaxin 1A (STX1A) can bind to VAMP2 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 VAMP2 and recombinant human STX1A. Briefly, STX1A was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to VAMP2-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. Measured by its binding ability in a functional ELISA. When recombinant VAMP2 is lmmobilized at 2 ug/mL(100 uLwell), the concentration of STX1A that produces 50% optimal bindingresponse is found to be approximately0.68ug/mL.