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|Antigen||Solute Carrier Family 1 (Glial High Affinity Glutamate Transporter), Member 3 (SLC1A3) Antibodies|
|Epitope||C-Term, AA 500-542 Alternatives|
|Reactivity||Human, Mouse (Murine), Rat (Rattus) Alternatives|
Immunocytochemistry (ICC), Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (IHC), ELISA, Western Blotting (WB)
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|Purification||Immunogen affinity purified|
|Immunogen||A synthetic peptide made to a C-terminal portion of the rat SLC1A3 protein (between residues 500-542) [UniProt P24942]|
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|Alternative Name||SLC1A3 / EAAT1 (SLC1A3 Antibody Abstract)|
|Background||Gene Symbol: SLC1A3|
|Molecular Weight||Theoretical MW: 60 kDa|
|Pathways||Sensory Perception of Sound, Synaptic Membrane, Dicarboxylic Acid Transport|
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|Application Notes||Western Blot 2 μg/mL, Flow Cytometry 1:200-1:500, ELISA 1:100-1:2000, Immunohistochemistry 1:10-1:500, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry-Paraffin 1:50-1:500, Immunohistochemistry-Frozen 1:10-1:500, Flow (Intracellular) 1:500The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
Western blot Protocol for SLC1A3 antibody
Western Blot Protocol:
1. Perform SDS-PAGE on protein samples to be analyzed, loading 10-40 µg of total protein per lane.
. Electro-blot the proteins to a suitable membrane (PVDF or Nitrocellulose) according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain the membrane with Ponceau S (or a similar product) to assess transfer success. Mark molecular weight standards where appropriate.
. Thoroughly rinse the membrane of stain with TBST.
. Incubate the membrane in blocking buffer (5 % non-fat milk in TBST or 5 % BSA in TBST) as appropriate, for 60 minutes.
. Dilute the SLC1A3 primary antibody as appropriate in blocking buffer and incubate for 60 minute at room temperature to overnight at 4 degrees C with gently shaking.
. Wash the membrane in TBST three times for 10 minutes each.
. Incubate the membrane in the appropriate secondary antibody prepared in blocking buffer (as per manufacturer's instructions) and incubate for 60 minutes at room temperature.
. Wash the membrane in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
. Incubate the membrane in the appropriate detection reagent in accordance with the manufacturer's instructions and image the blot.Note: Tween-20 can be added to the blocking, wash and antibody dilution buffers to a final concentration of 0.05-0.1 %.Immunohistochemistry-Paraffin protocol for SLC1A3 antibody Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 60 minutes at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4 degrees C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.
|Restrictions||For Research Use only|
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Buffer contains: 0.02 % Sodium Azide
|Precaution of Use||This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Handling Advice||Avoid freeze-thaw cycles|
|Storage||4 °C,-20 °C|
|Storage Comment||Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.|
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|Product cited in:||
Arnold, Salvatore: "Exercise-Mediated Increase in Nigral Tyrosine Hydroxylase Is Accompanied by Increased Nigral GFR-α1 and EAAC1 Expression in Aging Rats." in: ACS chemical neuroscience, Vol. 7, Issue 2, pp. 227-39, 2016 (Sample species: Rat (Rattus)). Further details: Western Blotting
Pinner, Tucholski, Haroutunian, McCullumsmith, Meador-Woodruff: "Decreased protein S-palmitoylation in dorsolateral prefrontal cortex in schizophrenia." in: Schizophrenia research, Vol. 177, Issue 1-3, pp. 78-87, 2016 (Sample species: Human). Further details: Western Blotting
Reyes-Aguirre, Lamas: "Oct4 Methylation-Mediated Silencing As an Epigenetic Barrier Preventing Müller Glia Dedifferentiation in a Murine Model of Retinal Injury." in: Frontiers in neuroscience, Vol. 10, pp. 523, 2016 (Sample species: Human).
Chotibut, Davis, Arnold, Frenchek, Gurwara, Bondada, Geddes, Salvatore: "Ceftriaxone increases glutamate uptake and reduces striatal tyrosine hydroxylase loss in 6-OHDA Parkinson's model." in: Molecular neurobiology, Vol. 49, Issue 3, pp. 1282-92, 2014 (Sample species: Rat (Rattus)). Further details: Western Blotting
Vollbrecht, Simmler, Blakely, Deutch: "Dopamine denervation of the prefrontal cortex increases expression of the astrocytic glutamate transporter GLT-1." in: Journal of neurochemistry, Vol. 130, Issue 1, pp. 109-14, 2014 (Sample species: Rat (Rattus)). Further details: Western Blotting
Hu, Takano, Xiang, Gilkes, Luo, Semenza: "Hypoxia-inducible factors enhance glutamate signaling in cancer cells." in: Oncotarget, Vol. 5, Issue 19, pp. 8853-68, 2014 (Sample species: Human). Further details: Western Blotting
Salvatore, Davis, Arnold, Chotibut: "Transient striatal GLT-1 blockade increases EAAC1 expression, glutamate reuptake, and decreases tyrosine hydroxylase phosphorylation at ser(19)." in: Experimental neurology, Vol. 234, Issue 2, pp. 428-36, 2012
Kobayashi, Millhorn: "Hypoxia regulates glutamate metabolism and membrane transport in rat PC12 cells." in: Journal of neurochemistry, Vol. 76, Issue 6, pp. 1935-48, 2001
Schmitt, Asan, Pueschel, Kugler: "Cellular and regional distribution of the glutamate transporter GLAST in the CNS of rats: nonradioactive in situ hybridization and comparative immunocytochemistry." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 17, Issue 1, pp. 1-10, 1997
Rothstein, Dykes-Hoberg, Pardo, Bristol, Jin, Kuncl, Kanai, Hediger, Wang, Schielke, Welty: "Knockout of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate." in: Neuron, Vol. 16, Issue 3, pp. 675-86, 1996
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