Aquaporin-5 Overexpression Lysate Summary
Description |
Aquaporin-5 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC206069
Accession#: NM_001651
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 |
AQP5 |
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 |
28.1 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 Aquaporin-5 Overexpression Lysate
Background
Aquaporins (AQPs) are a large family of integral membrane water transport channel proteins that facilitate the transport of water through the cell membrane. This function is conserved in animals, plants and bacteria. At least ten isoforms of aquaporin have been identified in mammals, designated AQP0 through AQP9. Aquaporins are widely distributed and it is not uncommon for more than one type of AQP to be present in the same cell. Although most aquaporins are only permeable to water, AQP3, AQP7 and AQP9 are permeable to urea and glycerol. AQP2 is the only water channel that is activated by vasopressin to enhance water reabsorption in the kidney collecting duct. Aquaporins are involved in renal water absorption, generation of pulmonary secretions, lacrimation, and the secretion and reabsorption of cerebrospinal fluid and aqueous humor. In the lung, the 27-kDa AQP5 is responsible for the majority of water transport across the apical membrane of type I alveolar epithelial cells. The gene encoding human AQP5 maps to chromosome 12q13.
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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