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Anti-GlyT2 (SLC6A5) (extracellular) Antibody

Na+/Cl--dependent glycine transporter 2

Cat #: AGT-012
Alternative Name Na+/Cl--dependent glycine transporter 2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: m, r
May also work in: h*
Immunogen
  • Peptide CVIGDHPKIQIKNS, corresponding to amino acid residues 333-346 of rat GlyT2 (Accession P58295). 2nd extracellular loop.
Accession (Uniprot) Number P58295
Gene ID 171148
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse - identical; human - 13/14 amino acid residues identical.
RRID AB_11121049.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ih, wb
May also work in: ic*, ifc*, ip*, lci*
Western blot
  • Western blot analysis of mouse brain (lanes 1 and 4), rat brain (lanes 2 and 5) and rat cerebellum (lanes 3 and 6) lysates:
    Western blot analysis of mouse brain (lanes 1 and 4), rat brain (lanes 2 and 5) and rat cerebellum (lanes 3 and 6) lysates:
    1-3. Anti-GlyT2 (SLC6A5) (extracellular) Antibody (#AGT-012), (1:200).
    4-6. Anti-GlyT2 (SLC6A5) (extracellular) Antibody, preincubated with GlyT2/SLC6A5 (extracellular) Blocking Peptide (#BLP-GT012).
Immunohistochemistry
  • Expression of glycine transporter 2 in mouse brain stem
    Expression of glycine transporter 2 in mouse brain stem
    Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using Anti-GlyT2 (SLC6A5) (extracellular) Antibody (#AGT-012), (1:200). A. GlyT2 (red) is expressed in neurons (arrow). B. Dapi staining of cell nuclei (blue) is used as a general cellular marker.
References
  1. Zafra, F. et al. (2008) IUBMB Life. 60, 810.
  2. Chen, N. et al. (2004) Pflugers. Arch. 447, 519.
  3. Supplisson, S. et al. (2002) FEBS Lett. 529, 93.
  4. Kristensen, A.S. et al. (2011) Pharmacol. Rev. 63, 585.
  5. Olivares, L. et al. (1995) J. Biol. Chem. 270, 9437.
  6. Massari, S. et al. (2005) J. Biol. Chem. 280, 7388.
  7. Liu, Q.R. et al. (1993) J. Biol. Chem. 268, 22802.
  8. Rees, M.I. et al. (2006) Nat. Genet. 38, 801.
Scientific background

Apart from its obvious biochemical functions, glycine is also an important inhibitory neurotransmitter. Following depolarization, glycine is released from synaptic vesicles, binds to glycine receptors (GlyRs) on postsynaptic membranes thereby causing hyperpolarization of postsynaptic neurons due to the massive influx of Cl- ions. Glycine is then taken up from the synaptic cleft via the glycine transporters GlyT1 and GlyT21.

GlyT1 and GlyT2 belong to the SLC6, Na+/Cl- dependent transporter family, of which members include transporters for GABA, serotonin, dopamine and norepinephrine2. Like all SLC6 members, GlyT1 and GlyT2 have 12 transmembrane domains and intracellular N- and C-terminals. Both can be found in different splice variants3,4. SLC6 transporters undergo post-translational modifications. For instance, GlyT1 and GlyT2 are glycosylated, which is important for their membrane trafficking5. Phosphorylation of these two transporters also takes place in a PKC-dependent manner, which may lead to down regulation of both transporters6. Pharmacologically, GlyT1 and GlyT2 can be differentiated by applying sarcosine which inhibits GlyT1 but not GlyT27.

GlyT1 and GlyT2 are broadly expressed in the nervous system; GlyT1 is concentrated in glial cells, while GlyT2 is present in glycenergic neurons in the spinal cord, brainstem and cerebellum1. GlyT1 can also be detected in the pancreas, uterus, stomach, spleen, liver and retina4.

GlyT1 has become a target for the treatment of schizophrenia, although a defect of the protein is not directly associated with the disorder. Inhibiting GlyT1 should lead to the increase in glutamatergic pathways, thereby decreasing psychotic effects in schizophrenic individuals. GlyT2 has been associated with hyperekplexia, a motor disorder characterized by neonatal hypertonia and startle reflex8.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 12/08/2021

Alomone Labs is pleased to offer a highly specific antibody directed against an extracellular epitope of the rat glycine transporter 2. Anti-GlyT2 (SLC6A5) (extracellular) Antibody (#AGT-012) can be used in western blot and immunohistochemistry analysis and has been designed to recognize GlyT2 from rat, mouse and human samples.

For research purposes only, not for human use
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