c-Fos Overexpression Lysate

Name: c-Fos Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-10794

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-10794

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for c-Fos Overexpression Lysate

c-Fos Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC202597

Accession#: NM_005252

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.

TMW

40.5 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 c-Fos Overexpression Lysate

Western Blot: c-Fos Overexpression Lysate [NBL1-10794]

Western Blot: c-Fos Overexpression Lysate (Adult Normal) [NBL1-10794] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for c-Fos.

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: c-Fos

c-Fos is an intermediate early gene that is one of four members of the FOS family of activator protein-1 (AP-1) transcription factors, which also includes Fra-1, Fra-2, and FosB (1-3). Under the FOS gene, human c-Fos is synthesized as a protein of 308 amino acids (aa) with a basic leucine zipper (bZip) domain important for dimerization, and a basic domain for interacting with DNA, with a theoretical molecular weight of ~40.6 kDa (4,5). c-Fos can heterodimerize with members of the JUN family (c-Jun, JunB, and JunD) to form the active transcription factor AP-1 complex (3,5). AP-1 related proteins bind TPA-related responsive elements (TRE), cAMP responsive elements (CRE), and related sequences, with c-Fos:c-Jun dimers partial to TRE sites (5).

In response to stimuli, c-Fos, which is encoded by protooncogenes, has a role in cell proliferation, differentiation, and transformation (3,6). A variety of stimuli can increase c-Fos expression such as growth factors, proinflammatory cytokines, UV radiation, neurotransmitters, hormones, injury, and stress (1,6). c-Fos has long been used as a marker for neuronal activity and is associated with neural and behavioral responses following stimuli (1-3, 6-7). Mouse studies have revealed that c-Fos is important for efficient neurogenesis and cortical development (3). Additionally, c-Fos signal can be used as a molecular marker for learning and memory, such as recognition and fear (2,7). Studies have found that repeated positive stimuli result in increased Fos expression while, conversely, repeated negative value stimuli are indicated by decreased signal (7). Intermediate early genes have also been implicated in neuropsychiatric disorders including showing altered c-Fos expression in a schizophrenia animal model (2). Furthermore, antipsychotics and antidepressants are both capable of impacting c-Fos expression (2).

References

1. Kovacs K. J. (1998). c-Fos as a transcription factor: a stressful (re)view from a functional map. Neurochemistry International. https://doi.org/10.1016/s0197-0186(98)00023-0

2. Gallo, F. T., Katche, C., Morici, J. F., Medina, J. H., & Weisstaub, N. V. (2018). Immediate Early Genes, Memory and Psychiatric Disorders: Focus on c-Fos, Egr1 and Arc. Frontiers in Behavioral Neuroscience. https://doi.org/10.3389/fnbeh.2018.00079

3. Velazquez, F. N., Caputto, B. L., & Boussin, F. D. (2015). c-Fos importance for brain development. Aging. https://doi.org/10.18632/aging.100862

4. Uniprot (P01100)

5. Wu, Z., Nicoll, M., & Ingham, R. J. (2021). AP-1 family transcription factors: a diverse family of proteins that regulate varied cellular activities in classical hodgkin lymphoma and ALK+ ALCL. Experimental Hematology & Oncology. https://doi.org/10.1186/s40164-020-00197-9

6. Shaulian, E., & Karin, M. (2001). AP-1 in cell proliferation and survival. Oncogene. https://doi.org/10.1038/sj.onc.1204383

7. Chung L. (2015). A Brief Introduction to the Transduction of Neural Activity into Fos Signal. Development & Reproduction. https://doi.org/10.12717/DR.2015.19.2.061

Long Name

FBJ/Finkel–Biskis–Jinkins Murine Osteosarcoma Viral Oncogene Homolog

Alternate Names

cFos, FOS, G0S7

Gene Symbol

FOS

Additional c-Fos Products

Product Documents for c-Fos Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for c-Fos 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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