FOXH1 (Forkhead Box H1, Human Homolog of Xenopus Forkhead Activin Signal Transducer 1)

FoxH1 (also known as FAST-1) is a 39kD, class 2, subfamily H member of the FOX \n(forkhead box) family of transcription factors. It is a nuclear protein that binds DNA promoter sites. It does not independently regulate transcription but interacts with SMAD2/4 to activate, and SMAD3/4 to inhibit gene transcription. Human FOXH1 is 365 amino acids (aa) in length. It contains an NLS (aa 22-30), one FKH (fork head) DNA-binding domain (aa 32-128) and a SMAD-interaction region (aa 273-354). Over aa 164-272, human FOXH1 is 55% identical to mouse FoxH1. \n\nApplications:\nSuitable for use in Western blot, Immunocytochemistry and Direct ELISA \n \nRecommended Dilutions: \nWestern blot: 0.1-0.2ug/ml. The detection limit is ~5 ng/lane under non-reducing and reducing conditions. \nImmunocytochemistr: 10ug/ml to detect human FoxH1 in human BG0IV stem cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at RT and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT. After blocking, cells were incubated with diluted primary antibody for 3 hours at RT followed by donkey anti-sheep lgG at room temperature for 1 hour. Between each step, cells were washed with PBS containing 0.1% BSA. \nDirect ELISA: 0.5-1.0ug/ml. The detection limit is~ 0.2 ng/well. \nOptimal dilutions should be determined by each laboratory for each application. \n\nHybridoma: \nProduced in sheep immunized with purified, E. coli-derived, recombinant human FoxH1 \n(rhFoxH1; aa 164-272; Accession # O75593). \n\nStorage:\nLyophilized samples are stable for twelve months from date of receipt when stored \nat -20 to -70 degrees C. Upon reconstitution, the antibody can be stored at 2-8 degrees C for 1 month without detectable loss of activity. Reconstituted antibody can also be aliquotted and stored frozen at -20 to -70 degrees C in a manual defrost freezer for six months without detectable loss of activity. Avoid repeated freeze-thaw cycles.

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