EAAT1/GLAST-1/SLC1A3 Overexpression Lysate Summary
Description |
EAAT1/GLAST-1/SLC1A3 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC230179
Accession#: NM_001166695
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
SLC1A3 |
Applications/Dilutions
Dilutions |
|
Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for EAAT1/GLAST-1/SLC1A3 Overexpression Lysate
Background
Human excitatory amino acid transporters (EAATs) are members of a family of high affinity sodium-dependent transporter molecules that regulate neurotransmitter concentrations at the excitatory glutamatergic synapses of the mammalian central nervous system. It is believed that these proteins reduce extracellular glutamate concentration, thereby modulating synaptic signaling. In addition, EAATs may also be important for the prevention of glutamate excitotoxicity. EAAT1 is prominently expressed in the frontal cortex, hippocampus and basal ganglia and is also reported to be found in heart, placenta, lung and striated muscle. EAAT1 shares some pharmacological similarities with EAAT3 and both are potent antagonists that appear to specifically block transport mediated by EAAT2. EAAT1 transports L-glutamate and also L- and D-aspartate, acting as a symport by co-transporting sodium. EAAT1 is essential for terminating the postsynaptic action of glutamate, by rapidly removing released glutamate from the synaptic cleft.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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