Bethyl Laboratories, Inc.
Antibodies Against Forkhead Box (FOX) Proteins
April 27, 2010
Overview


The first fork head (FKH) box (Fox) family member was discovered as a gene responsible for the "fork head" phenotype in D. melanogaster mutants. In these mutants, defects in FKH caused homeotic transformations that involved the replacement of foregut and hindgut by ectopic head structures resulting in two "spiked-head" structures in embryos (1). Since the discovery of FKH in flies, hundreds of Fox gene family members have been discovered in a range of species from yeast to humans. At least 43 members of the of the human Fox gene family have been identified (2); and based on phylogenetic analysis, 19 classes of Fox proteins have been established (FoxA-FoxS) (3). Fox proteins are defined by a highly conserved 100-residue forkhead DNA binding domain which consists of three alpha-helices, three-beta sheets, and two "wing" regions. The Fox proteins are DNA-binding proteins and generally function as transcriptional regulators; however duplication of the FKH domain through evolution has resulted in a family of proteins that exhibit a wide spectrum of functional diversity. The requirement of this family of proteins in processes ranging from organogenesis to speech acquisition exemplifies its importance through evolution, and along with its involvement in human disease, makes the Fox family a subject worthy of future study.
Selected Reviews
1. D. Weigel et al., "The Homeotic Gene Fork Head Encodes a Nuclear Protein and Is Expressed in the Terminal Regions of the Drosophila Embryo," Cell. 57, no. 4 (1989): 645-658.
2. M. Katoh and M. Katoh, "Human FOX Gene Family (Review)," Int.J Oncol. 25, no. 5 (2004): 1495-1500.
3. S. Hannenhalli and K. H. Kaestner, "The Evolution of Fox Genes and Their Role in Development and Disease," Nat Rev Genet. 10, no. 4 (2009): 233-240.

 



Detection of Human FOXK1 by WB and IP.

Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) and 293T (T; 50 mcg) cells.  Antibodies:  Affinity purified rabbit anti-FOXK1 antibody A301-728A used for WB at 0.04 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate. FoxK1 was also immunoprecipitated by rabbit anti-FOXK1 antibody A301-727A, which recognizes an upstream epitope. For blotting immunoprecipitated FOXK1, the ReliaBLOT Reagents and Procedures (Cat. No. WB120) were used. Detection:  Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).


Detection of Human FOXK2 by WB and IP.

Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) and 293T (T; 50 mcg cells.
Antibodies: Affinity purified rabbit anti-FOXK2 antibody A301-730A used for WB at 0.04 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate. FOXK2 was also immunoprecipitated by rabbit anti-FOXK2 antibody A301-729A, which recognizes an upstream epitope. For blotting immunoprecipitated FOXK2, the ReliaBLOT Reagents and Procedures (Cat. No. WB120) were used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).
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