Product Pathways - Nuclear Receptor Signaling
STF-1 (D1Z2A) XP® Rabbit mAb #12800
|12800S||100 µl (10 western blots)||---||In Stock||---|
|12800P||40 µl (4 western blots)||---||In Stock||---|
|12800||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat||Endogenous||50||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Bovine.
Specificity / Sensitivity
STF-1 (D1Z2A) XP® Rabbit mAb recognizes endogenous levels of total STF-1 protein. This antibody does not cross-react with LRH-1/NR5A2.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu184 of human STF-1 protein.
Western blot analysis of extracts from various cell lines using STF-1 (D1Z2A) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing DDK-tagged full-length human STF-1 (hSTF-1-DDK; +) or Myc/DDK-tagged full-length human LRH-1, isoform 2 (hLRH-1-Myc/DDK; +), using STF1 (D1Z2A) XP® Rabbit mAb (upper) and DYKDDDDK Tag Antibody #2368 (lower).
Immunoprecipitation of STF-1 from NCI-H295R cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or STF-1 (D1Z2A) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using STF-1 (D1Z2A) XP® Rabbit mAb.
Confocal immunofluorescent analysis of NCI-H295R (positive, left) and SW-13 (negative, right) cells using STF-1 (D1Z2A) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCI-H295R cells treated with dibutyryl cAMP (0.4mM) for 1h and either 10 µl of STF-1 (D1Z2A) XP® Rabbit mAb or 2 µl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human CYP17A1 intron 1 primers, SimpleChIP® Human StAR Intron 1 Primers #12864, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
The orphan nuclear receptor, steroidogenic factor 1 (STF-1, also called Ad4BP), is encoded by the NR5A1 gene and plays an instrumental role in directing the transcriptional control of steroidogenesis (1). Initially identified as a tissue-specific transcriptional regulator of cytochrome P450 steroid hydroxylases, research studies of both global (2) and tissue-specific knockout mice (3-6) have demonstrated that STF-1 is required for the development of adrenal glands, gonads, ventromedial hypothalamus, and for the proper functioning of pituitary gonadotropes. Indeed, humans with mutations that render STF-1 transcriptionally inactive can present with testicular failure, ovarian failure, and adrenal insufficiency (7,8). Furthermore, dysregulation of STF-1 has been linked to diseases such as endometriosis (9) and adrenocortical carcinoma (10).
Like other nuclear hormone receptors, STF-1 has a modular domain structure composed of an amino-terminal zinc finger DNA-binding domain, a ligand-binding domain, a carboxy-terminal AF-2 activation domain, and a hinge region with AF-1-like activation activity. STF-1 also contains a fushi tarazu factor 1 box, which functions as an accessory DNA binding domain (11). STF-1 is primarily phosphorylated at Ser203, which is thought to enhance its transcriptional activity by promoting complex formation with transcriptional cofactors (12). In addition to phosphorylation at Ser203, STF-1 is subject to SUMO conjugation and acetylation at ε-amino groups of target lysine residues. Whereas SUMOylation represses STF-1 function (13,14), acetylation enhances its transcriptional activity (15).
- Parker, K.L. and Schimmer, B.P. (1997) Endocr Rev 18, 361-77.
- Luo, X. et al. (1994) Cell 77, 481-90.
- Zhao, L. et al. (2001) Development 128, 147-54.
- Jeyasuria, P. et al. (2004) Mol Endocrinol 18, 1610-9.
- Pelusi, C. et al. (2008) Biol Reprod 79, 1074-83.
- Zhao, L. et al. (2008) Mol Endocrinol 22, 1403-15.
- Achermann, J.C. et al. (1999) Nat Genet 22, 125-6.
- Lourenço, D. et al. (2009) N Engl J Med 360, 1200-10.
- Bulun, S.E. et al. (2009) Mol Cell Endocrinol 300, 104-8.
- Figueiredo, B.C. et al. (2005) J Clin Endocrinol Metab 90, 615-9.
- Little, T.H. et al. (2006) Mol Endocrinol 20, 831-43.
- Hammer, G.D. et al. (1999) Mol Cell 3, 521-6.
- Chen, W.Y. et al. (2004) J Biol Chem 279, 38730-5.
- Lee, F.Y. et al. (2011) Dev Cell 21, 315-27.
- Chen, W.Y. et al. (2005) Mol Cell Biol 25, 10442-53.
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For Research Use Only. Not For Use In Diagnostic Procedures.
XP® is a trademark of Cell Signaling Technology, Inc.
SimpleChIP® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.