GFAP Overexpression Lysate

Novus Biologicals | Catalog # NBL1-11043

Novus Biologicals
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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.

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

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.

Type

Overexpression

Scientific Data Images for GFAP Overexpression Lysate

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

Glial fibrillary acidic protein (GFAP) is a class III intermediate filament protein that is largely expressed in astrocytes in addition to non-myelinating Schwann cells and glial cells (1,2). Other members of the type III intermediate filament family include desmin, peripherin, and vimentin (2-4). GFAP was first identified in the brains of multiple sclerosis patients (2). Human GFAP protein is 432 amino acids in length with a theoretical molecular weight of ~50 kDa (1,3). GFAP has at least 10 known isoforms, with the most prevalent and common in the brain being GFAP-alpha which is made of a head domain, a rod domain with four coils (1A, 1B, 2A, 2B) joined by linker regions, and a tail domain (1). GFAP is a marker of mature astrocytes, but is also expressed throughout development in both fetal and adult neural stem cells (2). While the exact function of GFAP is still elusive, it has been shown to play a role in cellular processes such as migration, mitosis, structural integrity, and signaling (2).

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

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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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