| VAChT

The Vesicular acetylcholine transporter (VAChT) is a neurotransmitter transporter which is responsible for loading acetylcholine (ACh) into secretory organelles in neurons making acetylcholine available for secretion. It is encoded by Solute carrier family 18, member 3 (SLC18A3) gene, located within the first intron of the choline acetyltransferase gene. VAChT is able to transport ACh into vesicles by relying on an exchange between protons (H+) that were previously pumped into the vesicle diffusing out, thus acting as an antiporter. ACh molecules are then carried into the vesicle by the action of exiting protons. Acetylcholine transport utilizes a proton gradient established by a vacuolar ATPase.
 

| Typical data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, wash technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data are provided below for reference only.

 

Concentration (ng/mL)	20	10	5	2.5	1.25	0.625	0.312	0
OD	2.14	1.63	1.16	0.78	0.57	0.36	0.24	0.07
Corrected OD	2.07	1.56	1.09	0.71	0.5	0.29	0.17	-

 

| Repeatability

Intra-assay Precision (Precision within an assay)：CV% < 10%; Inter-assay Precision (Precision between assays)：CV% < 10%
 

| Recovery

The recovery of Human VAChT spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

 

Sample Type	Range(%)	Average Recovery(%)
Tissue Homogenates(n=8)	83-101	96
Cell Lysates(n=8)	92-105	101

 

| Linearity

Samples were spiked with high concentrations of Human VAChT and diluted with Universal Diluent to produce samples with values within the range of the assay.

 

		Tissue Homogenates(n=4)	Cell Lysates(n=4)
1 : 2	Range (%)	84-95	88-99
	Average (%)	91	93
1 : 4	Range (%)	89-103	87-108
	Average (%)	95	98