Western blot analysis was performed on whole cell extracts of PC-12 (Lane 1). The blot was probed with Anti-EAAC1 (Glutamate Transporter) Mouse Monoclonal Antibody (Product # 32-1000, 2 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 ug/ml, 1:2500 dilution). A 57 kDa band corresponding to EAAC1 was observed in the cell line tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12% Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Rat|
|Published species reactivity||Rat, Human|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Synthetic peptide corresponding to amino acids 161-177 of the rat glutamate transporter EAAC1 sequence.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||1-5 ug|
|Western Blot (WB)||2-4 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
L-glutamate is the major excitatory neurotransmitter in mammalian central nervous systems. To maintain glutamate levels below excitoxic levels, excess glutamate at excitatory synaptic clefts is removed by ion-coupled glutamate transporters. Four glutamate transporters have been identified: the sodium-dependent GLAST-1, GLT-1, EAAC-1 and the chloride-dependent EAAT-4. EAAC-1 is expressed in the CNS and also found in kidney and small intestine. Structural features of glutamate transporters are believed to include eight membrane-spanning alpha-helices and a loop-pore structure which is unique among secondary transporters but may resemble loop-pores found in ion channels. A second distinctive structural feature is the presence of a highly amphipathic membrane-spanning helix that provides a hydrophilic path through the membrane. Mice with homozygous deletions of the EAAC-1 transporter gene develop dicarboxylic aminoaciduria and display reduced spontaneous locomotor activity although no neurodegeneration was observed over a period of 12 months.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Rat||1:500||Impaired spinal cord glutamate transport capacity and reduced sensitivity to riluzole in a transgenic superoxide dismutase mutant rat model of amyotrophic lateral sclerosis.||Dunlop J,Beal McIlvain H,She Y,Howland DS||The Journal of neuroscience : the official journal of the Society for Neuroscience (23:1688)||2003|
|Immunohistochemical localization of the neuron-specific glutamate transporter EAAC1 (EAAT3) in rat brain and spinal cord revealed by a novel monoclonal antibody.||Shashidharan P,Huntley GW,Murray JM,Buku A,Moran T,Walsh MJ,Morrison JH,Plaitakis A||Brain research (773:139)||1997|