TAK1 Overexpression Lysate Summary
Description |
TAK1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC204454
Accession#: NM_003188
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 |
MAP3K7 |
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 |
64 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 TAK1 Overexpression Lysate
Background
TAK1 (or MAP3K7) was shown to participate in regulation of transcription by transforming growth factor beta (TGF beta). TAK1 is stimulated in response to TGF beta and bone morphogenetic protein. These results suggest that TAK1 functions as a mediator in the signaling pathway of TGF beta superfamily members. TAB1 and TAB2 are TAK1 binding proteins that may function as activators of the TAK1 (TGF b activated kinase 1) MAPKKK in TGF b signal transduction. TAB1 induced TAK1 activation promoted the dissociation of active forms of IKKa and IKK b from active TAK1, whereas the IKK mutants remained to interact with active TAK1. TNF a activated endogenous TAK1, and the kinase negative TAK1 acted as a dominant negative inhibitor against TNF a induced NFkB activation. TAK1 was suggested to act as a regulatory kinase of IKKs.
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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