Glutamate Transporter EAAT4 (GTRAP41, Excitatory Amino Acid Transporter 4)

For Research Use Only. Not for use in diagnostic procedures.

Chromosome: 20; NC_006602.2 (50137157..50147431)

Polyclonal

IgG

Host: Rabbit
Source: Rat

The rat peptide is 86% conserved in human GTRAP41/beta-spectrin 3. 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.

Purified by immunoaffinity chromatography.

Affinity Purified
Supplied as a liquid in PBS, pH 7.4 and 0.1% sodium azide, 40% glycerol.

May be stored at 4 degree 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 degree C. Aliquots are stable for at least 12 months at -20 degree 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.

Small volumes of anti-SLC1A6 antibody vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

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 two proteins, designated GTRAP41 and GTRAP48 (for glutamate transporter EAAT4 associated protein) that specifically interact with the intracellular carboxy-terminal domain of EAAT4 have been cloned and characterized. The proteins are proposed to modulate glutamate transport activity. GTRAP41 is 2388 aa protein (~270kD). It has seventeen 16-amino acid spectrin repeats, 2 alpha-actinin domains, and a pleckstrin homology domain. GTRAP48 is 1527 aa (~170kD). GTRAP48 possesses a PDZ domain, a regulatory G protein-signaling sequence, tandem dbl homology and pleckstrin homology domains characteristic of guanine nucleotide exchange factors for the Rho family of G proteins, and 2 proline-rich sequences. brain tissue. GTRAP41 and GTRAP48 are mainly expressed in the brain, and lower levels were also detected in liver and kidney. EAAT4, GTRAP41, and GTRAP48 are all immunolocalized in cerebellar Purkinje cell soma and dendrites, with little axonal staining. GTRAP48 was found to activate Rho. Expression of GTRAP48 induced the reorganization of the actin cytoskeleton. When GTRAP41 and GTRAP48 are coexpressed, GTRAP41 colocalized with actin in structures that resemble actin-stress fibers, a typical Rho-dependent effect. Overexpression of GTRAP41 and GTRAP48 enhanced glutamate uptake. GTRAP41 and GTRAP48 may therefore enhance glutamate transport either through an increase in the catalytic rate of the transporter or through an increase in cell-surface availability or by stabilizing EAAT4 at the membrane.

Abs to Proteins

ELISA, Western Blot

Suitable for use in ELISA, Western Blotting. Other applications not tested.
Dilution: Western Blot: 1:1000-1:5000 for antiserum and 1-10ug/ml for affinity pure antibody using Chemiluminescence technique. GTRAP41 is ~270kD protein (1).
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.

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??1. Jackson M et al (2001) Nature 410, 89-93; Ohara O et al (1998) Brain Res. Mol. Brain res. 57, 181; Stanekewich MC et al (1998) PNAS 95, 14158; Rumenapp U eat al (1999) FEBS Lett. 459, 313; Fukuhara JBC 274, 5868-5879

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