|Calculated MW||56537 Da|
|Homology||Mouse - identical.|
|Other Names||Vesicular acetylcholine transporter, VAChT, rVAT, Solute carrier family 18 member 3, Slc18a3, Vacht, Vat|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)DAVRLREVQGKDGGE, corresponding to amino acid residues 495-509 of rat vesicular acetylcholine transporter (Accession Q62666). Cytoplasmic, C-terminus.Not recommended for human samples.|
|Peptide Confirmation||Confirmed by mass-spectrography and amino acid analysis.|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.8 mg/ml.|
|Buffer After Reconstitution||Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.|
|Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Storage After Reconstitution||The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).|
|Control Antigen Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Control Antigen Reconstitution||100 µl DDW.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||2 µg peptide per 1 µg antibody.|
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Provided below are standard protocols that you may find useful for product applications.
In cholinergic neurons, acetylcholine is synthesized from choline and acetyl-coenzyme A (acetyl-CoA). Vesicular acetylcholine transporter (VAChT) transports acetylcholine from the cytoplasm to the lumen of synaptic vesicles where following depolarization, acetylcholine is released in the synaptic cleft and activates muscarinic and nicotinic receptors located on postsynaptic membranes1-3. Acetylcholine released in the synaptic cleft is rapidly hydrolyzed into choline and acetate. Since choline is not de novo synthesized and thereby only made available through diet uptake, choline is recycled back into the cells in order to regenerate acetylcholine1,2. VAChT is a member of the SLC18 family of transporters. It has 12 membrane spanning domains, with extracellular N- and C-terminal tails. The transporter exchanges cytoplasmic acetylcholine for two vesicular protons4-6. VAChT is a selective marker of cholinergic neurons and could be used therefore to determine the loss of cholinergic neurons in Alzheimer’s or Alzheimer’s related disease7. VAChT knockout mice die after birth. They display a lack of stimulated release of acetylcholine and an underdeveloped neuromuscular junction4. Abgent is pleased to offer a highly specific antibody directed against an epitope of rat VAChT. Anti-Vesicular Acetylcholine Transporter antibody (#AG1291) can be used in western blot analysis. It has been designed to recognize VAChT from mouse and rat samples.
References 1. Ribeiro, F.M. et al. (2006) J. neurochem. 97, 1. 2. Okuda, T. and Haga, T. (2003) Neurochem. Res. 28, 483. 3. Bravo, D.T. et al. (2004) J. Neurochem. 91, 766. 4. de Castro, B.M. et al. (2009) Mol. Cell. Biol. 29, 5238. 5. Nguyen, M.L. et al. (2000) Biochemistry 37, 13400. 6. Parsons, S.M. (2000) FASEB J. 14, 2423. 7. Masson, J. et al. (1999) Pharmacol. Rev. 51, 439.
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