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EAAT2 antibody extracellular domain - 250 203 K.O.

EAATs are transmembrane proteins involved in the removal of extracellular glutamate
Rabbit polyclonal purified antibody
Cat. No.: 250 203
Amount: 50 µg
Price: $380.00
Cat. No. 250 203 50 µg specific antibody, lyophilized. Affinity purified with the immunogen. Albumin and azide were added for stabilization. For reconstitution add 50 µl H2O to get a 1mg/ml solution in PBS. Then aliquot and store at -20°C to -80°C until use.
Antibodies should be stored at +4°C when still lyophilized. Do not freeze!
Applications
 
WB: 1 : 1000 (AP staining) gallery  
IP: not tested yet
ICC: yes (see remarks) gallery  
IHC: 1 : 1000 up to 1 : 2000 (see remarks) gallery  
IHC-P: 1 : 500 (see remarks) gallery  

Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section.

Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section.

Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section.

Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section.

Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.

Immunogen Synthetic peptide corresponding to AA 146 to 161 from mouse EAAT2 (UniProt Id: P43006)
Reactivity Reacts with: rat (P31596), mouse (P43006).
Other species not tested yet.
Specificity K.O. validated
Matching control protein/peptide 250-2P
Remarks

ICC: This antibody yields excellent results on mouse cells, but is not recommended for rat samples, as it produces non-specific nuclear staining in those specimens.
IHC: This antibody yields excellent results on mouse tissue but is not recommended for rat samples, as it produces non-specific nuclear staining in those specimens.

Fixation with 4% PFA and 0,1% glutaraldehyde according to Danbolt et al. 1998 is recommended.
IHC-P: This antibody yields excellent results on mouse tissue but is not recommended for rat samples, as it produces non-specific nuclear staining in those specimens.

Data sheet 250_203.pdf

References for EAAT2 - 250 203

Rapid recycling of glutamate transporters on the astroglial surface.
Michaluk P, Heller JP, Rusakov DA
eLife (2021) 10: . 250 203 WB; tested species: mouse
Astrocyte dysfunction increases cortical dendritic excitability and promotes cranial pain in familial migraine.
Romanos J, Benke D, Pietrobon D, Zeilhofer HU, Santello M
Science advances (2020) 623: eaaz1584. 250 203 WB; tested species: mouse
Differences in glutamate uptake between cortical regions impact neuronal NMDA receptor activation.
Romanos J, Benke D, Saab AS, Zeilhofer HU, Santello M
Communications biology (2019) 2: 127. 250 203 WB; tested species: mouse
Chronic Toxoplasma infection is associated with distinct alterations in the synaptic protein composition.
Lang D, Schott BH, van Ham M, Morton L, Kulikovskaja L, Herrera-Molina R, Pielot R, Klawonn F, Montag D, Jänsch L, Gundelfinger ED, et al.
Journal of neuroinflammation (2018) 151: 216. 250 203 WB, IHC; tested species: mouse
Strategies for immunohistochemical protein localization using antibodies: What did we learn from neurotransmitter transporters in glial cells and neurons.
Danbolt NC, Zhou Y, Furness DN, Holmseth S
Glia (2016) 6412: 2045-2064. 250 203 WB
Region-specific changes in gene expression are associated with cognitive deficits in the alpha-synuclein-induced model of Parkinson's disease: A transcriptomic profiling study.
Manchinu MF, Pala M, Palmas MF, Diana MA, Maschio A, Etzi M, Pisanu A, Diana FI, Marongiu J, Mansueto S, Carboni E, et al.
Experimental neurology (2024) 372: 114651. 250 203 IHC; tested species: rat
Crym-positive striatal astrocytes gate perseverative behaviour.
Ollivier M, Soto JS, Linker KE, Moye SL, Jami-Alahmadi Y, Jones AE, Divakaruni AS, Kawaguchi R, Wohlschlegel JA, Khakh BS
Nature (2024) : . 250 203 IHC; tested species: mouse
Glutamate transporters EAAT2 and EAAT5 differentially shape synaptic transmission from rod bipolar cell terminals.
Tang FS, Yuan HL, Liu JB, Zhang G, Chen SY, Ke JB
eNeuro (2022) : . 250 203 IHC; tested species: mouse
Glioblastoma hijacks neuronal mechanisms for brain invasion.
Venkataramani V, Yang Y, Schubert MC, Reyhan E, Tetzlaff SK, Wißmann N, Botz M, Soyka SJ, Beretta CA, Pramatarov RL, Fankhauser L, et al.
Cell (2022) : . 250 203 IHC; tested species: mouse
Chronic Toxoplasma infection is associated with distinct alterations in the synaptic protein composition.
Lang D, Schott BH, van Ham M, Morton L, Kulikovskaja L, Herrera-Molina R, Pielot R, Klawonn F, Montag D, Jänsch L, Gundelfinger ED, et al.
Journal of neuroinflammation (2018) 151: 216. 250 203 WB, IHC; tested species: mouse
Characterization of Tissue Plasminogen Activator Expression and Trafficking in the Adult Murine Brain.
Stevenson TK, Lawrence DA
eNeuro () 54: . 250 203 IHC; tested species: mouse
Pentylenetetrazole-induced Seizure Susceptibility in the Tau58/4 Transgenic Mouse Model of Tauopathy.
Van Erum J, Valkenburg F, Van Dam D, Paul De Deyn P
Neuroscience (2019) : . 250 203 IHC-P; tested species: mouse
Cat. No.: 250 203
Amount: 50 µg
Price: $380.00
Rapid recycling of glutamate transporters on the astroglial surface.
Michaluk P, Heller JP, Rusakov DA
eLife (2021) 10: . 250 203 WB; tested species: mouse
Astrocyte dysfunction increases cortical dendritic excitability and promotes cranial pain in familial migraine.
Romanos J, Benke D, Pietrobon D, Zeilhofer HU, Santello M
Science advances (2020) 623: eaaz1584. 250 203 WB; tested species: mouse
Differences in glutamate uptake between cortical regions impact neuronal NMDA receptor activation.
Romanos J, Benke D, Saab AS, Zeilhofer HU, Santello M
Communications biology (2019) 2: 127. 250 203 WB; tested species: mouse
Chronic Toxoplasma infection is associated with distinct alterations in the synaptic protein composition.
Lang D, Schott BH, van Ham M, Morton L, Kulikovskaja L, Herrera-Molina R, Pielot R, Klawonn F, Montag D, Jänsch L, Gundelfinger ED, et al.
Journal of neuroinflammation (2018) 151: 216. 250 203 WB, IHC; tested species: mouse
Strategies for immunohistochemical protein localization using antibodies: What did we learn from neurotransmitter transporters in glial cells and neurons.
Danbolt NC, Zhou Y, Furness DN, Holmseth S
Glia (2016) 6412: 2045-2064. 250 203 WB
Region-specific changes in gene expression are associated with cognitive deficits in the alpha-synuclein-induced model of Parkinson's disease: A transcriptomic profiling study.
Manchinu MF, Pala M, Palmas MF, Diana MA, Maschio A, Etzi M, Pisanu A, Diana FI, Marongiu J, Mansueto S, Carboni E, et al.
Experimental neurology (2024) 372: 114651. 250 203 IHC; tested species: rat
Crym-positive striatal astrocytes gate perseverative behaviour.
Ollivier M, Soto JS, Linker KE, Moye SL, Jami-Alahmadi Y, Jones AE, Divakaruni AS, Kawaguchi R, Wohlschlegel JA, Khakh BS
Nature (2024) : . 250 203 IHC; tested species: mouse
Glutamate transporters EAAT2 and EAAT5 differentially shape synaptic transmission from rod bipolar cell terminals.
Tang FS, Yuan HL, Liu JB, Zhang G, Chen SY, Ke JB
eNeuro (2022) : . 250 203 IHC; tested species: mouse
Glioblastoma hijacks neuronal mechanisms for brain invasion.
Venkataramani V, Yang Y, Schubert MC, Reyhan E, Tetzlaff SK, Wißmann N, Botz M, Soyka SJ, Beretta CA, Pramatarov RL, Fankhauser L, et al.
Cell (2022) : . 250 203 IHC; tested species: mouse
Chronic Toxoplasma infection is associated with distinct alterations in the synaptic protein composition.
Lang D, Schott BH, van Ham M, Morton L, Kulikovskaja L, Herrera-Molina R, Pielot R, Klawonn F, Montag D, Jänsch L, Gundelfinger ED, et al.
Journal of neuroinflammation (2018) 151: 216. 250 203 WB, IHC; tested species: mouse
Characterization of Tissue Plasminogen Activator Expression and Trafficking in the Adult Murine Brain.
Stevenson TK, Lawrence DA
eNeuro () 54: . 250 203 IHC; tested species: mouse
Pentylenetetrazole-induced Seizure Susceptibility in the Tau58/4 Transgenic Mouse Model of Tauopathy.
Van Erum J, Valkenburg F, Van Dam D, Paul De Deyn P
Neuroscience (2019) : . 250 203 IHC-P; tested species: mouse
Background

Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system. After the release of glutamate from synaptic vesicles into the synaptic cleft during neurotransmission, excitatory amino acid transporters (EAATs) remove extracellular glutamate to avoid excitotoxic levels (1).
Five EAATs with differential expression patterns have been described so far: EAAT1, also referred to as GLAST and SLC1A3, has neuroprotective potential following ischemia and occurs in reactive astrocytes and activated microglia. EAAT2 (GLT-1, SLC1A2) is the most abundant isoform and is primarily expressed in astrocytes. Both variants show high levels in brain and retina. EAAT3 / SLC1A1, EAAT4 / SLC1A6 and EAAT5 / SLC1A7 are expressed in neurons (2). EAAT4 shows weak expression in the forebrain and high levels in the cerebellum, where it mainly locates to Purkinje cells (3). EAAT5 primarily occurs in the retina, where it locates very close to glutamate release sites. In K.O. mice flicker resolution is considerably compromised (4). Recent findings suggest that EAAT5 is an abundant isoform, expressed also in non-neuronal peripheral tissues (5).