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.

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

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.

Array NBL1-10794

• c-Fos Antibodies
• c-Fos ELISA Kits
• c-Fos Lysate
• c-Fos Proteins