Revision 1

#37342Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

65

Source/Isotype:

Rabbit IgG

UniProt ID:

#P30531

Entrez-Gene Id:

6529

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Frozen) 1:100 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

GAT1 (E7J1B) Rabbit mAb recognizes endogenous levels of total GAT1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human GAT1 protein.

Background

The solute carrier 6 gene (SLC6), also known as the neurotransmitter–sodium-symporter family or Na+/Cl- -dependent transporter, encodes for proteins that regulate neurotransmitter (NTT) transport, including monoamine transmitters serotonin, dopamine, and norepinephrin (SERT), GABA transmitters (GAT1, GAT2, GAT3, and BGT1), and glycine transmitters (GLYT1 and GLYT2) (1). These proteins express ubiquitously in the brain and regulate the release and uptake of neurotransmitters in terminal clefts, in both neuronal and non-neuronal cells (2-4). Dysregulation of NTT-transporters is associated with neurological disease like epilepsy, schizophrenia, anxiety, bipolar disorder, and addictions to cocaine and methamphetamines (1). Inhibitors of NTT-transporters are widely used as drugs to treat disorders like depression (tricyclic antidepressants), and antiepileptic tiagabine (5). GAT1 is the only GABA transporter genetically studied in GAT1-KO mouse models where an accumulation of extracellular GABA decreased anxiety and depression-like behaviors (6-8). The lack or reduction of GAT1 diminished aggression in mice, and a condition known as hypoalgesia, where there is a decreased sensitivity to painful stimuli (8,9). GAT1 posttranslational modifications include phosphorylation at Tyr107 (IL1), and Tyr317 (IL3), and these mutations identify as the phospho-acceptor-sites, therefore regulating GAT1 (10,11). GABA trafficking is regulated by Tyr phosphorylation, and it has been shown that activation of adenosine A2A receptors in the hippocampus synaptosomes enhanced BAGA uptake by opposing a constitutive PKC-mediated downregulation of GAT1 (11-13).

  1. Kristensen, A.S. et al. (2011) Pharmacol Rev 63, 585-640.
  2. Borden, L.A. (1996) Neurochem Int 29, 335-56.
  3. Moldavan, M. et al. (2017) J Neurophysiol 118, 3092-106.
  4. Lorenz-Guertin, J.M. and Jacob, T.C. (2018) Dev Neurobiol 78, 238-70.
  5. Skovstrup, S. et al. (2010) ChemMedChem 5, 986-1000.
  6. Bragina, L. et al. (2008) J Neurochem 105, 1781-93.
  7. Jensen, K. et al. (2003) J Neurophysiol 90, 2690-701.
  8. Liu, G.X. et al. (2007) Neuropsychopharmacology 32, 1531-9.
  9. Xu, Y.F. et al. (2008) J Neurosci Res 86, 465-70.
  10. Whitworth, T.L. and Quick, M.W. (2001) J Biol Chem 276, 42932-7.
  11. Law, R.M. et al. (2000) J Biol Chem 275, 23986-91.
  12. Giros, B. et al. (1996) Nature 379, 606-12.
  13. Chen, N. et al. (2004) J Biol Chem 279, 5508-19.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#37342

GAT1 (E7J1B) Rabbit mAb

Western Blotting Image 1: GAT1 (E7J1B) Rabbit mAb Expand Image
Western blot analysis of extracts from various tissues using GAT1 (E7J1B) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Immunoprecipitation Image 1: GAT1 (E7J1B) Rabbit mAb Expand Image
Immunoprecipitation of GAT1 protein from mouse brain lysates. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GAT1 (E7J1B) Rabbit mAb. Western blot analysis was performed using GAT1 (E7J1B) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunofluorescence Image 1: GAT1 (E7J1B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of rat cerebellum using GAT1 (E7J1B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: GAT1 (E7J1B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of rat retina using GAT1 (E7J1B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 3: GAT1 (E7J1B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of rat hippocampus (left) and striatum (right) using GAT1 (E7J1B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).