F2H Kit Androgen Receptor

Description
Analyze and manipulate the intramolecular interaction between the N- and C-terminal domains of Androgen Receptor (AR) with ChromoTek Fluorescent-Two Hybrid (F2H®) AR.
Wild-type (wt), as well as several mutant AR forms can be investigated.

Specificity
AR is expressed as two separate domains
Bait:    AR ligand binding domain (AR-LBD) fused to GFP
Prey:   AR N-terminal domain (AR-NTD) fused to RFP
Bait and prey interact.

Applications
Detect early steps of Androgen Receptor activation
Real-time agonists and antagonists screening
Visualized in live mammalian cells

Available AR variants
AR-LBD: wt, W741L, F876L, T877A, F876L-T877A

Click on the left picture for a live cell analysis example
Live-cell F2H imaging of reversible interactions between AR-LBD-GFP (green) and AR-NTD-RFP (red) in F2H-BHK cells

ProductSizeCodePriceBuy
ProductF2H®-Kit AR-N/C wt (Androgen Receptor, wt)SizeF2H®-BHK Cell line: 5x106 cells, Platform-ARLBD(wt)-ARNTD: 100 µg, Positive Control: 30 µgCodef2hk-ar-wtPrice $ 1385
Buy+
ProductPlatform-ARLBD(wt)-ARNTDSize100 µgCodef2h-ar-wtPrice $ 525
Buy+
ProductPlatform-ARLBD(W741L)-ARNTDSize100 µgCodef2h-ar-w741lPrice $ 525
Buy+
ProductPlatform-ARLBD(F876L)-ARNTDSize100 µgCodef2h-ar-f876lPrice $ 525
Buy+
ProductPlatform-ARLBD(T877A)-ARNTDSize100 µgCodef2h-ar-t877aPrice $ 525
Buy+
ProductPlatform-ARLBD(F876L/T877A)-ARNTDSize100 µgCodef2h-ar-f876l/t877aPrice $ 525
Buy+
ProductPositive ControlSize30 µgCodef2h-caPrice $ 120
Buy+

Cell line
BHK (baby hamster kidney)

Detection
Epifluorescence microscope

Benefits

  • Analyze modulators of the human Androgen Receptor
  • Detect early steps of Androgen Receptor activation
  • Visualized in live mammalian cells
  • Real-time agonists and antagonists screening
  • Convenient readout using an epi-fluorescence microscope

Please inquire if you want to learn more about our F2H Assay Kits.

Principle

  • AR is expressed as two separate domains: AR-LBD (ligand binding domain), comprising the AR’s C-terminus, either wt or mut, fused to GFP: AR-LBD-GFP
  • AR-NTD (N-terminal domain), comprising the AR’s N-terminus, fused to RFP: AR-NTD-RFP
  • F2H BHK cells express components of the protein-protein interaction platform, which recruits GFP-tagged AR-LBD to a specific location in the nucleus
  • AR-LBD-GFP and AR-NTD-RFP constructs are co-transfected into the F2H BHK cells
  • AR-LBD-GFP forms a bright green spot at the specific location in the nuclei whereas AR-NTD-RFP is diffusely distributed in the nucleus
  • Upon stimulation by an agonist (e.g. DHT), immobilized AR-LBD-GFP undergoes a conformation change and recruits AR-NTD-RFP: AR-NTD-RFP forms a bright red spot superimposed with the green spot.

Results

Green spot is present, red spot is absent => assay works: no interaction
Green spot is present, red spot is present at the same location => assay works: interaction

Reversible screening:
The F2H AR assay is fully reversible: the AR wt interaction can be induced by an agonist (e.g. 0.25 nM DHT) and then subsequently disrupted by addition of a potent antagonist (e.g. 10 µM bicalutamide).

Please click on picture at the top of the page to properly see the animation.

About Androgen Receptor

The human Androgen receptor (AR) is an important target for developing drug therapies combating prostate cancer. The very early event in AR activation, namely the intramolecular interaction between the N- and C-terminus of AR, is very important.

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Only for research applications, not for diagnostic or therapeutic use!