Adenosine Human Transporter Cell Based Antagonist Uptake LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Adenosine Na⁺-independent nucleoside transporters (Equilibrative nucleoside transporters, ENTs) are integral membrane proteins that mediate the bidirectional, passive transport of nucleosides and nucleobases across cellular membranes, driven by concentration gradients. ENTs play essential roles in nucleotide metabolism by enabling the recycling of purine and pyrimidine nucleosides for DNA synthesis, energy metabolism, and cellular signaling. In humans, four ENT isoforms (ENT1–ENT4) have been identified, each with distinct substrate specificities, tissue distribution patterns, and physiological functions. ENT1 and ENT2 are the most well-characterized, widely expressed, and primarily involved in transporting naturally occurring nucleosides and nucleobases. Beyond their physiological roles, ENTs are pharmacologically important for mediating the uptake of nucleoside-derived drugs, including anticancer and antiviral agents.
Family:
Transporter
Sub Family:
SLC29 / Nucleoside
Class:
SLC
Accession Number:
XM_011514341
Organism:
Human
Construct Details:
Endogenous
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human histocytic lymphoma U937 cells (ATCC, CRL-1593) are used. The cells (1 x 10⁶/well) are preincubated with test compound and/or vehicle in modified Tris buffer pH 7.4 at room temperature for 20 minutes, and 10 nM [3H]Adenosine is then added for an additional 2-minute incubation period. Reduction of [3H]Adenosine uptake by 50 percent or more (≥50%) relative to 1 μM NBTI response indicates significant inhibitory activity. If significant inhibition of uptake is observed, item #315550-1 is suggested as a follow-up assay to evaluate for possible compound-induced cytotoxicity.
Incubation:
2 min at RT
Control Inhibitor:
NBTI
Criteria For Significance:
≥50% Inhibition of [3H]Adenosine uptake relative toNBTI response
Test Concentration / Dose:
Compounds are screened at 10, 3, 1, 0.3, 0.1, 0.03, 0.01 and 0.003 μM
Test Sample Requirements:
75 μl of 10 mM stock or 1 mg (pre-weighed) powder (assuming FW ≦500 and purity 100%)
Minimum Order Quantity:
16
Turnaround Time
Standard:
20 Business Days
Reference Compound Data
Clinical Relevance
Adverse / Beneficial Action:
Blockade of adenosine uptake may also prolong its accumulation in the heart to provide protection against the deleterious effects of ischemia reperfusion.
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
Questions? Contact Us
You can easily get in touch with a Eurofins Discovery Expert or get help from our Customer Service team. Reach out with questions, or to discuss your specific discovery challenges.
