GFAP Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-11043
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for GFAP Overexpression Lysate
GFAP Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC204548
Accession#: NM_002055
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.
Expression Host: HEK293T
Plasmid: RC204548
Accession#: NM_002055
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.
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.
TMW
49.7 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.
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
Overexpression
Scientific Data Images for GFAP Overexpression Lysate
![Western Blot: GFAP Overexpression Lysate [NBL1-11043]](https://resources.bio-techne.com/images/products/GFAP-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-11043-img0002.jpg "Western Blot: GFAP Overexpression Lysate [NBL1-11043]")
Western Blot: GFAP Overexpression Lysate [NBL1-11043]
Western Blot: GFAP Overexpression Lysate (Adult Normal) [NBL1-11043] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GFAP.Formulation, Preparation, and Storage
Formulation
RIPA buffer
Concentration
The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -80C. Avoid freeze-thaw cycles.
Background: GFAP
An increase in GFAP levels is often associated with neuroinflammation which results in the activation and proliferation of astroglia cell population (1,2). GFAP expression is also observed in brains of patients with neurodegenerative diseases including Alzheimer's and Parkinson's, epilepsy disorders, and brain injuries (1-4). Lesion sites associated with neurodegeneration can exhibit an array of gliosis characteristics from glial scarring with reduced astrocyte proliferation to activated, GFAP-positive astrocytes surrounding amyloid plaques (2). Furthermore, the GFAP gene is a target of single nucleotide polymorphisms in the coding region, considered a gain-of-function mutation, characterized by astrocytic inclusions, termed Rosenthal fibers, resulting in Alexander Disease (1-4). GFAP is also a center of many post-translational modifications, such as phosphorylation, which can alter various aspects of filament assembly (1,4).
References
1. Yang, Z., & Wang, K. K. (2015). Glial fibrillary acidic protein: from intermediate filament assembly and gliosis to neurobiomarker. Trends in Neurosciences. https://doi.org/10.1016/j.tins.2015.04.003
2. Hol, E. M., & Capetanaki, Y. (2017). Type III Intermediate Filaments Desmin, Glial Fibrillary Acidic Protein (GFAP), Vimentin, and Peripherin. Cold Spring Harbor Perspectives in Biology. https://doi.org/10.1101/cshperspect.a021642
3. Potokar, M., Morita, M., Wiche, G., & Jorgacevski, J. (2020). The Diversity of Intermediate Filaments in Astrocytes. Cells. https://doi.org/10.3390/cells9071604
4. Viedma-Poyatos, a., Pajares, M. A., & Perez-Sala, D. (2020). Type III intermediate filaments as targets and effectors of electrophiles and oxidants. Redox Biology. https://doi.org/10.1016/j.redox.2020.101582
Long Name
Glial Fibrillary Acidic Protein
Alternate Names
ALXDRD, FLJ45472, GFAP, GFAP astrocytes, glial fibrillary acidic protein
Gene Symbol
GFAP
Additional GFAP Products
Product Documents for GFAP Overexpression Lysate
Product Specific Notices for GFAP 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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