Glt 1

Manufactured by Merck Group

Sourced in United States

GLT-1 is a laboratory equipment product developed by Merck Group. It serves as a glutamate transporter that helps regulate the levels of glutamate, a key neurotransmitter, in research settings.

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Lab products found in correlation

Glial fibrillary acidic protein (gfap), by Merck Group (2 mentions) Glast, by Merck Group (2 mentions) Fluorchem fc2 image system, by Bio-Techne (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Tdp 43, by Proteintech (2 mentions) Gapdh, by Merck Group (1 mentions) Neuronal nuclei (neun), by Merck Group (1 mentions) Pierce bca assay, by Thermo Fisher Scientific (1 mentions) Protease and phosphatase inhibitor, by Merck Group (1 mentions) Glast, by Abcam (1 mentions) Eclipse 80i, by Nikon (1 mentions) Goat anti mouse igg cy3 antibody, by Jackson ImmunoResearch (1 mentions) Goat anti rabbit igg cy2, by Jackson ImmunoResearch (1 mentions) Glua1, by Merck Group (1 mentions) Glua2, by Merck Group (1 mentions) Hrp conjugated secondary antiserum, by Jackson ImmunoResearch (1 mentions)

Spelling variants of this product

Guinea pig anti-GLT-1 (7 citations) Anti-GLT-1 (4 citations)

7 protocols using glt 1

Western Blot Protein Extraction and Analysis

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Proteins were extracted by homogenizing samples in lysis buffer (1% sodium dodecyl sulfate (SDS), 100 mM Tris(hydroxymethyl)aminomethane (Tris) buffer, pH 7.5, supplemented with protease and phosphatase inhibitors (Sigma)), followed by two rounds of sonication for seven seconds. Lysates were centrifuged for 5 minutes at 16,000xg, followed by ThermoScientific’s Pierce BCA assay to determine protein concentration.
Proteins were heated to 60°C for 15 minutes with 2x loading buffer (100 mM Tris, pH 6.8, 4% SDS, in Laemmli-sodium dodecyl sulfate, 600 mM B-mercaptoethanol, 200 mM Dithiothreitol (DTT), and 20% glycerol). Equal amounts of protein per sample were loaded on a 4–20% gradient precast SDS polyacrylamide gel, followed by transfer to a PVDF membrane at 100V for one hour. Primary antibodies used were as follows: Aquaporin 4 (Alomone, rabbit, polyclonal, 1:2000); Gapdh (Millipore, chicken, polyclonal, 1:2000); Gfap (Millipore, mouse, polyclonal, 1:30,000); Glast (Abcam, rabbit, polyclonal, 1:500); and Glt-1 (Millipore, chicken, polyclonal, 1:10,000); Cd11b (Alamone, rabbit, polyclonal, 1:500); Map2 (Sigma, mouse, monoclonal, 1:500); NeuN (Millipore, rabbit, polyclonal, 1:500). All HRP secondary antibodies were used at a 1:2,000 concentration.

Holt L.M, & Olsen M.L. (2016). Novel Applications of Magnetic Cell Sorting to Analyze Cell-Type Specific Gene and Protein Expression in the Central Nervous System. PLoS ONE, 11(2), e0150290.

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Hippocampal Protein Expression Analysis

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Protein samples from the hippocampus of the different groups were separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. Nonspecific binding sites were blocked by immersing the membranes in 5% bovine serum albumin in PBS at room temperature, followed by incubation with primary antibodies. Subsequently, the membranes were incubated with appropriate secondary antibodies (Santa Cruz, USA). All secondary antibodies were horseradish peroxidase conjugated. ECL Western Blotting Substrate (Pierce, USA) was used to detect the immunoreactive signals with an ECL-based Fluorchem^® FC2 image system (Alpha Innotech, USA). Primary antibodies included GLAST, GLT-1 and GS antibody (Millipore, USA). All western blot analyses were performed in triplicate. The FluorChem FC2 software was used to analyze the gray value of the protein expression in each group.

Guan R.L., Wang T., Li J., Zhao F., Zheng G., Shen X.F., Aschner M., Du K.J, & Liu M.C. (2022). Effects of Co-exposure to Lead and Manganese on Learning and Memory Deficits. Journal of environmental sciences (China), 121, 65-76.

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Immunohistochemical Analysis of Rat Brain

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Anesthetized rats were transcardially perfused with saline followed by 4% paraformaldehyde (PFA). After post-fixation (overnight, at 4 °C) in 4% PFA, the cryoprotected tissue was sliced (at 30-μm) using a microtome (Microm HM550, Walldorf, German). Primary antibodies against NeuN (1:1000, Millipore, Billerica, MA), GLT-1 (1:1000, Millipore; 1:1000, Abcam, Cambridge, MA), S100 (1:1000, sigma), and GFAP (1:500, Millipore) were used. Goat anti-rabbit IgG Cy2, and Goat anti-mouse IgG Cy3 antibodies (Jackson Immuno-Research, West Grove, PA) were used at 1:400 dilutions. Control experiments showed no interaction between secondary antibodies, showed species specificity for secondary antibodies, and showed no background staining by secondary antibodies. Immuno-positive cells were visualized with a CCD camera connected to a Nikon Eclipse 80i (Nikon, Melville, NY). The LHb region was identified as described (Aizawa et al., 2012 (link)).

Kang S., Li J., Bekker A, & Ye J.H. (2017). Rescue of glutamate transport in the lateral habenula alleviates depression- and anxiety-like behaviors in ethanol-withdrawn rats. Neuropharmacology, 129, 47-56.

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Western Blot Analysis of Hippocampal Proteins

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Protein samples from the hippocampus of the different groups were separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. Nonspecific binding sites were blocked by immersing the membranes in 5% bovine serum albumin in PBS at room temperature, followed by incubation with primary antibodies. Subsequently, the membranes were incubated with appropriate secondary antibodies (Santa Cruz, USA). All secondary antibodies were horseradish peroxidase conjugated. ECL Western Blotting Substrate (Pierce, USA) was used to detect the immunoreactive signals with an ECL-based Fluorchem® FC2 image system (Alpha Innotech, USA). Primary antibodies included GLAST, GLT-1 and GS antibody (Millipore, USA). All western blot analyses were performed in triplicate. The FluorChem FC2 software was used to analyze the gray value of the protein expression in each group.

Guan R., Wang T., Dong X., Du K., Li J., Zhao F., Xu J., Li B., Zheng G., Shen X., Cao B., Wang J., Aschner M., Liu M., & Chen R. (2021). Effects of Co-exposure to Lead and Manganese on Learning and Memory Deficits.

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Subcellular Fractionation and Immunoblotting

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The PFC and NAc regions were dissected, homogenized in sucrose buffer containing protease inhibitors, and spun at 1000 g. The supernatant was retrieved and then spun at 10000 g. The resulting pellet was re-suspended in sucrose buffer and protease inhibitors, and spun at 10000 g. The supernatant was then discarded and the pellet containing the membrane fraction was suspended in 1% sodium dodecyl sulfate (SDS) in RIPA. Proteins were separated using 10% SDS-PAGE and transferred to PVDF membrane. The membranes were probed overnight at 4°C with primary antibodies diluted in 5% milk/Tris-buffered saline with 0.1% Tween 20. GLT-1 (1:5000, Chemicon, Billerica, MA) was measured in both the PFC and NAc, and GluA1 and GluA2 (1:1000, Chemicon, Billerica, MA) were measured in the NAc only. After incubation with HRP-conjugated secondary antiserum (Jackson Immuno; 1:10,000), immunoreactive bands on the membranes were detected by ECL Plus. Band density was measured using NIH Image J software.

LaCrosse A.L., Hill K, & Knackstedt L.A. (2015). Ceftriaxone attenuates cocaine relapse after abstinence through modulation of nucleus accumbens AMPA subunit expression. European neuropsychopharmacology : the journal of the European College of Neuropsychopharmacology, 26(2), 186-194.

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Intrathecal Infusion and Spinal Cord Analysis

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Thirty days after the start of intrathecal infusions, the animals were anesthetized with isoflurane, and perfused transcardially with saline, followed by 4% paraformaldehyde (PFA) for 10 min. The lumbar enlargements of spinal cords were dissected and post-fixed in 4% PFA for 24 h and removed to 30% sucrose/PBS for another 2 days. Serial 20-μm frozen sections were cut from the middle of the lumbar enlargement for ~ 5 mm in the caudal direction.
After every 10 sections, 3–5 serial sections were set aside for staining. Immunohistochemical stains were carried out on floating sections, blocked (10% HS, 1 h), and exposed to primary antibody in 10% HS, 0.3% Triton-X 100 (SMI-32, 1:8000 IP, Sternberger Monoclonals, Berkeley, CA; GLT-1, 1:1000 IF, Chemicon, Temecula, CA; 3-nitrotyrosine, 10 μg/ml IF, Upstate Biotechnology, Waltham, MA; TDP-43, 1:10,000 IP, Proteintech Group, Chicago, IL). Labeling was visualized either by routine ABC immunoperoxidase techniques or under fluorescence using secondary antibodies linked to fluorophores (Alexafluor 488, Molecular Probes, Eugene, OR).

Yin H.Z., Yu S., Hsu C.I., Liu J., Acab A., Wu R., Tao A., Chiang B.J, & Weiss J.H. (2014). Intrathecal infusion of BMAA induces selective motor neuron damage and astrogliosis in the ventral horn of the spinal cord. Experimental neurology, 0, 1-9.

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BMAA Immunohistochemistry Protocol

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BMAA was obtained from Sigma (St. Louis, MO). Antibodies were from the following sources: SMI-32, Sternberger Monoclonals, Berkeley, CA; GLT-1, Chemicon, Temecula, CA; 3-nitrotyrosine, Upstate Biotechnology, Waltham, MA; TDP-43, Proteintech Group, Chicago, IL. For fluorescence labeling, we used secondary antibodies linked to Alexafluor 488 (Molecular Probes, Eugene, OR). All other chemicals and reagents were obtained from common commercial sources.

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