EAAT3 Antibody (Rabbit mAb) [A15N21]

Application: Reactivity: Mouse, Rat

Usage Information

Dilution
WB1:1000 IP1:30 IHC1:1000 IF1:100

Application
WB, IP, IHC, IF

Reactivity
Mouse, Rat

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
57 kDa 150-250 kDa,50-75 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Datasheet & SDS

Biological Description

Specificity
EAAT3 Antibody (Rabbit mAb) [A15N21] detects endogenous levels of total EAAT3 protein.

Clone
A15N21

Synonym(s)
Eaac1, Eaat3, Slc1a1, Excitatory amino acid transporter 3, Excitatory amino-acid carrier 1, Sodium-dependent glutamate/aspartate transporter 3, Solute carrier family 1 member 1

Background
Excitatory amino acid transporter 3 (EAAT3/SLC1A1) is a high‑affinity, Na⁺‑dependent glutamate and aspartate transporter of the EAAT family that resides predominantly on neuronal membranes, especially dendrites and axon terminals, where it regulates local excitatory amino acid concentrations and terminates postsynaptic glutamatergic signaling. EAAT3 shares the general EAAT architecture of multiple transmembrane helices forming a substrate-binding pocket coupled to Na⁺ and H⁺ co‑transport and K⁺ counter‑transport, and exhibits a distinctive pattern in which a substantial fraction of the protein is retained in intracellular compartments, with rapid trafficking to and from the plasma membrane controlled by neuronal activity and signaling pathways such as α‑protein kinase C and PI3K. At excitatory synapses, EAAT3 is positioned postsynaptically, complementing astrocytic EAAT1/GLAST and EAAT2/GLT‑1, and its transport activity lowers glutamate near postsynaptic receptors, buffering NMDA and AMPA receptor activation, shaping synaptic transmission and plasticity, and preventing excessive receptor stimulation that could lead to excitotoxic damage. EAAT3 also transports cysteine and serves as the major neuronal cysteine uptake route; this function makes EAAT3 the rate‑limiting factor for glutathione synthesis in EAAT3‑expressing neurons, so transporter activity directly supports antioxidant capacity and protection against oxidative stress, while in GABAergic neurons EAAT3‑mediated glutamate uptake supplies precursor for GABA synthesis and thus contributes to inhibitory neurotransmission. Regulation of EAAT3 occurs at multiple levels: transcriptional control adjusts expression, intracellular signaling modulates transporter trafficking between cytoplasm and surface, and astrocyte-secreted factors such as cholesterol influence its membrane localization, producing dynamic changes in glutamate and cysteine uptake in response to synaptic activity and metabolic state. Loss-of-function mutations in SLC1A1 cause dicarboxylic aminoaciduria with elevated urinary glutamate and aspartate, reflecting its role in renal and neuronal reuptake of anionic amino acids, and EAAT3 knockout mice show reduced neuronal glutathione, increased oxidative stress and age-dependent loss of substantia nigra neurons, identifying EAAT3 as essential for long-term dopaminergic neuron survival. Genetic variation in SLC1A1 has been associated with obsessive-compulsive disorder, and clinical plus preclinical data support involvement of EAAT3 dysregulation in OCD-related circuit function, while experimental models of Parkinson’s disease demonstrate that EAAT3-mediated glutamate uptake and metabolic use via glutamate dehydrogenase are critical gateways for neuroprotection, with knockdown of EAAT3 abolishing glutamate’s beneficial effects on ATP production and oxidative damage in dopaminergic neurons.

Background

Excitatory amino acid transporter 3 (EAAT3/SLC1A1) is a high‑affinity, Na⁺‑dependent glutamate and aspartate transporter of the EAAT family that resides predominantly on neuronal membranes, especially dendrites and axon terminals, where it regulates local excitatory amino acid concentrations and terminates postsynaptic glutamatergic signaling. EAAT3 shares the general EAAT architecture of multiple transmembrane helices forming a substrate-binding pocket coupled to Na⁺ and H⁺ co‑transport and K⁺ counter‑transport, and exhibits a distinctive pattern in which a substantial fraction of the protein is retained in intracellular compartments, with rapid trafficking to and from the plasma membrane controlled by neuronal activity and signaling pathways such as α‑protein kinase C and PI3K. At excitatory synapses, EAAT3 is positioned postsynaptically, complementing astrocytic EAAT1/GLAST and EAAT2/GLT‑1, and its transport activity lowers glutamate near postsynaptic receptors, buffering NMDA and AMPA receptor activation, shaping synaptic transmission and plasticity, and preventing excessive receptor stimulation that could lead to excitotoxic damage. EAAT3 also transports cysteine and serves as the major neuronal cysteine uptake route; this function makes EAAT3 the rate‑limiting factor for glutathione synthesis in EAAT3‑expressing neurons, so transporter activity directly supports antioxidant capacity and protection against oxidative stress, while in GABAergic neurons EAAT3‑mediated glutamate uptake supplies precursor for GABA synthesis and thus contributes to inhibitory neurotransmission. Regulation of EAAT3 occurs at multiple levels: transcriptional control adjusts expression, intracellular signaling modulates transporter trafficking between cytoplasm and surface, and astrocyte-secreted factors such as cholesterol influence its membrane localization, producing dynamic changes in glutamate and cysteine uptake in response to synaptic activity and metabolic state. Loss-of-function mutations in SLC1A1 cause dicarboxylic aminoaciduria with elevated urinary glutamate and aspartate, reflecting its role in renal and neuronal reuptake of anionic amino acids, and EAAT3 knockout mice show reduced neuronal glutathione, increased oxidative stress and age-dependent loss of substantia nigra neurons, identifying EAAT3 as essential for long-term dopaminergic neuron survival. Genetic variation in SLC1A1 has been associated with obsessive-compulsive disorder, and clinical plus preclinical data support involvement of EAAT3 dysregulation in OCD-related circuit function, while experimental models of Parkinson’s disease demonstrate that EAAT3-mediated glutamate uptake and metabolic use via glutamate dehydrogenase are critical gateways for neuroprotection, with knockdown of EAAT3 abolishing glutamate’s beneficial effects on ATP production and oxidative damage in dopaminergic neurons.

References
https://pubmed.ncbi.nlm.nih.gov/16800850/ https://pubmed.ncbi.nlm.nih.gov/29800605/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%