G-protein beta-subunit-like (GBL), also known as MTOR Associated Protein, LST8 Homolog (mLST8), is a 326 amino acid (aa) protein that is comprised of seven WD40 repeats and has a predicted molecular weight of approximately 36 kDa. The human protein shares 97% aa sequence identity with the mouse and rat orthologs. GBL binds the kinase domain of TOR and has been shown to stimulate its activity. GBL is found in both complexes formed by TOR, TORC1 and TORC2. It is required for TORC1-dependent phosphorylation of p70 S6 Kinase and IRS1 following stimulation with serum and insulin, and TORC2-dependent phosphorylation of Akt on Ser473 in response to serum, Insulin, and IFN-beta. GBL also negatively regulates TNF-alpha-induced NFkB activation via binding to IKK alpha and IKK beta. The physiological importance of GBL is highlighted by the fact that knockout mice display embryonic lethality, possibly due to defective vascular development. Additionally, female mice harboring heterozygous null mutations for both TOR and GBL have an increased life span, suggesting that GBL may also be involved in the aging process.
GBL Overexpression Lysate
Novus Biologicals | Catalog # NBL1-10995
Key Product Details
Species
Applications
Product Summary for GBL Overexpression Lysate
GBL Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC210610
Accession#: NM_022372
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.
Product Specifications
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
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.
Type
Scientific Data Images for GBL Overexpression Lysate
Western Blot: GBL Overexpression Lysate [NBL1-10995]
Western Blot: G protein beta subunit like Overexpression Lysate (Adult Normal) [NBL1-10995] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for G protein beta subunit like.Formulation, Preparation, and Storage
Formulation
Concentration
Shipping
Storage
Background: GBL
Long Name
Alternate Names
Gene Symbol
Additional GBL Products
Product Documents for GBL Overexpression Lysate
Product Specific Notices for GBL Overexpression Lysate
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.
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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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars