SV2A Overexpression Lysate Summary
| Description |
SV2A Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC208322
Accession#: NM_014849
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 |
SV2A |
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. |
| Theoretical MW |
82.5 kDa. Disclaimer note: The 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. |
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 SV2A Overexpression Lysate
Background
SV2A (Synaptic Vesicle protein 2A) is an integral membrane glycoproteins present in synaptic vesicles. SV2s have 12 transmembrane domains predicted by sequence analysis. There are three characterized isoforms, SV2A, SV2B and SV2C. SV2A is expressed ubiquitously throughout the brain. SV2B has a more restricted distribution with varying degrees of coexpression with SV2A. SV2C is more closely related to SV2A but shows a very restricted expression pattern. The highest expression levels were observed in phylogenetically old brain areas like pallidum, the midbrain and the olfactory bulb. SV2A is probably involved in the maintenance of a pool of synaptic vesicles competent for calcium-stimulated exocytosis. SV2A is a highly glycosylated synaptic vesicle protein with homology to transmembrane transporters.
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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