Glutamate Transporter EAAT4 (Excitatory Amino Acid Transporter 4)

Antibodies / Antibodies-Proteins

-20°C

Supplied as a liquid in PBS, pH 7.4 and 0.09% sodium azide, 40% glycerol.

A 21AA synthetic peptide near the cytoplasmic, C-terminus of rat EAAT4 (1) was synthesized, conjugated to KLH.

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. Glutamate transporters (525-573aa) display about 55% homology and are predicted to contain up to 6-10 transmembrane domains. Immunolocalization studies indicate that GLT1 is localized in astroglial cells throughout the brain and spinal cord. EAAC1 is specific for certain neurons and purkinje cells, and specifically enriched in cortex, hippocampus, and caudate-putamen and confined to presynaptic and postsynaptic elements. GLAST has been observed in both neuron and astroglia. It is most abundant in Bergmann glia, cortex, hippocampus and cerebellum. EAAT4 has properties of ligand gated Cl-channel. It is localized mainly in cerebellar Purkinje cells in rat and human CNS. EAAT5 has only been cloned from human. It is primarily expressed in retina.

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 antibody using Chemiluminescence technique (2). EAAT4 monomers in freshly made rat brain extracts appeared as fuzzy ~65kD protein (2). High MW bands may be see after oxidation (2).

ELISA: 1:10, 000-50, 000 for neat serum and 0.5-1ug/ml for affinity pure. Control peptide can be used to coat ELISA plates at 1ug/ml.

Optimal dilutions to be determined by the researcher.

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 5-10ug of control peptide per 1ug of affinity pure IgG or 1ul of antiserum) to demonstrate antibody specificity.

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.