Connexin 32/GJB1 Overexpression Lysate Summary
Description |
Connexin 32/GJB1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC205109
Accession#: NM_000166
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 |
GJB1 |
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 (cat# 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 |
31.8 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 Connexin 32/GJB1 Overexpression Lysate
Background
Gap junctions are aggregations of intercellular channels that directly connect the cytoplasm of adjacent cells. Gap junctions coordinate cellular and organ function in tissues and are involved in metabolic cooperation between cells, synchronization of cellular physiological activities, growth control and developmental regulation. The gap junction channels allow intercellular exchange of ions, nucleotides and small molecules between adjacent cells. Unlike other membrane channels, intercellular channels span two plasma membranes and require the contribution of hemichannels, called connexons, from both participating cells. These channels are permeate to molecules as large as 1 kD, and they have been detected in virtually every cell type in mammals, except mature skeletal muscle, spermatozoa and erythrocytes. Two connexons interact in the extracellular space to form the complete intercellular channel. Each connexon is composed of six similar or identical proteins, which are termed connexins. Connexins (Cx) are a multi-gene family of highly related proteins ranging between molecular weights of 26 and 70 kD. At least a dozen distinct connexin genes have been identified and many are expressed in a tissue-specific manner. Two distinct lineages have been identified in mammals. One termed class I or beta group to which Cx26, Cx30, Cx31, Cx31.1 and Cx32 belong, and the other termed class II or alpha group, represented by Cx33, Cx37, Cx40, Cx43 and Cx46.
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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