Competitive fluorescent adenosine receptor antagonist (apparent KD values are 7.50, 7.37 and 7.30 for A2A, A3 and A1 respectively). Antagonizes the activity of NECA, an adenosine receptor agonist. Inhibits cAMP accumulation and stimulates inositol phosphate accumulation (pKb values are 6.4 and 6.5 respectively). Exhibits no intrinsic agonist activity.
Figure 1. A2A-SPAP cells assayed against NECA and 1 µM HB7814
Figure 2. A3-SPAP cells assayed against NECA and 1 µM HB7814
Fluorescence imaging with HB7814
HB7814(30 nM) binding to live CHO cells expressing adenosine A1 receptors. Binding blocked by unlabelled competitor XAC (10 µM). Nuclei counter-stained with Hoechst.
Figure 1. A2A-SPAP cells assayed against NECA and 1 µM HB7814
Figure 2. A3-SPAP cells assayed against NECA and 1 µM HB7814
Fluorescence imaging with HB7814
HB7814(30 nM) binding to live CHO cells expressing adenosine A1 receptors. Binding blocked by unlabelled competitor XAC (10 µM). Nuclei counter-stained with Hoechst.
For ligand binding; fluorescence imaging; high content analysis; kinetic analysis; cell sorting at adenosine A1 / A2A / A3 receptors use solutions up to 100 nM.
Pharmacological validation
The CellAura fluorescent adenosine antagonist [XAC] ligand was shown to antagonize the activity of the adenosine receptor agonist adenosine-5'-N-ethyluronamide (NECA), in three separate recombinant CHO cell lines expressing the human A1, A2A or A3 receptor and a cyclic AMP-responsive secreted placental alkaline phosphatase (SPAP) reporter gene.The cyclic AMP-induced expression of SPAP was measured under basal and forskolin-stimulated (maximal) conditions. Addition of CellAura fluorescent adenosine antagonist [XAC] to the basal or forskolin-stimulated cells did not significantly alter basal and stimulated SPAP levels, demonstrating that CellAura fluorescent adenosine antagonist [XAC] has no intrinsic agonist activity.To determine the apparent KD for CellAura fluorescent adenosine antagonist [XAC], cells were treated with varying concentrations of NECA alone, or in the presence of 1µM CellAura fluorescent adenosine antagonist [XAC], and the cyclic AMP-induced expression of SPAP measured.The apparent KD at A1, A2A and A3 receptors was calculated from the rightward shift of the agonist response curve in the presence of CellAura fluorescent adenosine antagonist [XAC], compared to the response curve for the agonist alone, for each receptor-expressing cell line.
Solubility & Handling
Storage instructions
-20°C (protect from light)
Solubility overview
Soluble in DMSO
Handling
After thawing individual aliquots for use, we recommend briefly sonicating the sample to ensure it is fully dissolved and the solution is homogeneous. We do not recommend using the product after subjecting it to repetitive freeze-thaw cycles.
Shipping conditions
The product, supplied in a dry form, is stable at ambient temperature for periods of up to a few days and does not require shipping on ice/dry ice.
Important
This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use.
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