Glutamate is the main excitatory neurotransmitter in the brain.
To date five glutamate Transporters have been cloned: GLAST (EAAT1), GLT1 (EAAT2), EAAC1 (EAAT3), EAAT4, and EAAT5.
These transporters are believed to be critical in reducing potentially toxic extracellular concentration of glutamate by rapid uptake into nerve terminals and glial cells.
Most recently, vesicular type transporters for glutamate, termed VGLUTs (VGLUT1/BNPI, VGLUT2/DNPI, and EAT-4), have been cloned and characterized that are related to phosphate transporters.
Although neurons exhibit Na-dependent Pi transport, the biological role of Pi uptake is not clear.
Proximal tubules in the kidney reabsorb Pi in the glomeruli by the action of a group of phosphate transporters (Type 1-NaPi related, type 2-NaPi-2 related, and type 3-viral receptor-related.
These receptors show weak (~20% identity) between various subtypes.
A distinct type of brain specific Na+-dependent phosphate (Pi) transporter (BNPI), originally characterized as a plasma membrane transporter has been localized in a subset of glutamatergic neurons (amygdla,cereberal cortex, and hippocmaplus) and identified as VGLUT1 (rat and human 560 aa; ~60kD, ~30% homology with type-1 Pi-transporters & ~75-80% homology with VGLUT2).
Interestingly, a sequence induced by subtoxic levels of NMDA in cerebelar granules also belongs to this family of transporters.
BNPI/VGLUT1 is expression is restricted to the brain, where it is predominantly located in synaptic vesicles.
VGLUT1 shows strong sequence homology (~48%) to EAT-4, a C.
elegans protein that appears to have specific presynaptioc role in glutamtergic transmission.
Applications:Suitable for use in ELISA, Western Blotting.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000-1:5000 for neat serum and 1-10ug/ml for affinity pure using ECL.
A band of 95-110kD has been detected in brush border membrane (2)
ELISA:1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity purified.
Control peptide can be used to coat ELISA plates at 1ug/ml.
Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are stable for at least 12 months at -20°C.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
仕様
Size:100ul
Host:chicken
Source Antibody:rat
Grade:Serum
Purity:Serum
Form:Supplied as a liquid, neat serum, 40% glycerol.
Specificity:The rat peptide is 93% conserved in human VGLUT1. No significant sequence homology is observed with VGLUT2, EAT-4, or other NaPi-related (Type-1 to Type-III) transporters. Antibody crossreactivity in various species is not established. Control peptide, because of its low MW (<3kD), is not suitable for Western. It is recommed for ELISA, dot blot, or for antibody blocking experiments (use 5-10 ug of control peptide per 1 ug of aff pure IgG or 1 ul of antiserum) to demonstrate antibody specificity.
Isotype:IgY
Calc Applications Abbrev:E WB
Calc Crossreactivity:Rt
Immunogen:A 19 AA synthetic peptide witin the C-terminus of rat VGLUT1 was synthesized, conjugated to KLH