VAChT/SLC18A3 Overexpression Lysate

Western Blot: Vesicular Acetylcholine Transporter Overexpression Lysate (Adult Normal) [NBP2-10859] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Vesicular Acetylcholine Transporter.

• Reactivity: Hu
• Applications: WB

VAChT/SLC18A3 Overexpression Lysate Summary

• Description:

  VAChT/SLC18A3 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC202886
  
  Accession#: NM_003055
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: SLC18A3

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 56.7 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for VAChT/SLC18A3 Overexpression Lysate

• MGC12716
• SLC18A3
• solute carrier family 18 (vesicular acetylcholine), member 3
• Solute carrier family 18 member 3
• solute carrier family 18, member 3
• VAChT
• VACHTvesicular acetylcholine transporter

Background

Vesicular Acetylcholine Transporter (VAChT), (approx. 70kD protein), belongs to the family of vesicular monoamine transporters(VMATs), which include VMAT1 and VMAT2 and the C.elegans putative ACh transporter unc-17. Members of this family function to concentrate neurotransmitters into synaptic vesicles through exchange of protons for neurotransmitters. VAChT is a functional transporter for the neurotransmitter acetylcholine(ACh). ACh is synthesized in the cytoplasm by choline acetyl transferase (ChAT) and transported by VAChT into synaptic vesicles where it is stored until released. After release from presynaptic nerve terminals ACh is hydrolyzed by extracellular ACh-esterases(AChE) to choline and acetate. VAChT mRNA is expressed in all known major cholinergic neurons in the central and peripheral nervous system. VAChT is abundantly expressed in the CNS and is mainly localized in small synaptic vesicles in cholinergic nerve terminals. VAChT provides a specific marker for cholinergic neurons for the study of cholinergic transmission in experimental models, in Alzheimer's disease and other nervous system disorders.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: SLC18A3