Revision 5

#12800Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13285

Entrez-Gene Id:

2516

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:100
Chromatin IP 1:50
Chromatin IP-seq 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology

Bovine

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu184 of human STF-1 protein.

Background

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).

  1. Parker, K.L. and Schimmer, B.P. (1997) Endocr Rev 18, 361-77.
  2. Luo, X. et al. (1994) Cell 77, 481-90.
  3. Zhao, L. et al. (2001) Development 128, 147-54.
  4. Jeyasuria, P. et al. (2004) Mol Endocrinol 18, 1610-9.
  5. Pelusi, C. et al. (2008) Biol Reprod 79, 1074-83.
  6. Zhao, L. et al. (2008) Mol Endocrinol 22, 1403-15.
  7. Achermann, J.C. et al. (1999) Nat Genet 22, 125-6.
  8. Lourenço, D. et al. (2009) N Engl J Med 360, 1200-10.
  9. Bulun, S.E. et al. (2009) Mol Cell Endocrinol 300, 104-8.
  10. Figueiredo, B.C. et al. (2005) J Clin Endocrinol Metab 90, 615-9.
  11. Little, T.H. et al. (2006) Mol Endocrinol 20, 831-43.
  12. Hammer, G.D. et al. (1999) Mol Cell 3, 521-6.
  13. Chen, W.Y. et al. (2004) J Biol Chem 279, 38730-5.
  14. Lee, F.Y. et al. (2011) Dev Cell 21, 315-27.
  15. Chen, W.Y. et al. (2005) Mol Cell Biol 25, 10442-53.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 5
#12800

STF-1 (D1Z2A) XP® Rabbit mAb

Western Blotting Image 1: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
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 Blotting Image 2: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
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 Image 1: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
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.
Immunofluorescence Image 1: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
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 Immunoprecipitation Image 1: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H295R cells treated with dibutyryl cAMP (0.4mM) for 1h and STF-1 (D1Z2A) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across STAR, a known target gene of STF-1 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H295R cells treated with dibutyryl cAMP (0.4mM) for 1h and STF-1 (D1Z2A) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 8 (upper), including STAR (lower), a known target gene of STF-1 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: STF-1 (D1Z2A) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H295R cells treated with dibutyryl cAMP (0.4mM) for 1h and eitherSTF-1 (D1Z2A) XP® Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human CYP17A1 intron 1 primers, Simple ChIP® 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.