Tubulin Rat [3H] Colchicine Binding LeadHunter Assay - TW

Item: 219100

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OVERVIEW

DOCUMENTS/REFERENCES

Biological Information

Background Information:

Colchicine has a relatively selective anti-inflammatory effect in acute gouty arthritis and has also been used prophylactically for this disorder. Binding to tubulin, colchicine interferes with the function of mitotic spindles and causes depolymerization and disappearance of the fibrillar microtubules in granulocytes and other motile cells. This may be the basis for inhibition of migration of granulocytes into the inflamed area. Further, colchicine binding to tubulin inhibits its polymerization, unlike taxanes which stabilize microtubules, inhibits mitosis in metaphase due to failure of spindle formation. Thus, colchicine-like compounds may be therapeutically useful for the treatment of gouty arthritis and certain tumors. Inhibition of rapidly proliferating gastrointestinal epithelial cells may cause nausea, emesis, diarrhea and abdominal pain.

Family:

Cytoskeleton

Sub Family:

Tubulin

Organism:

Rat

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Binding

Detection Method:

Radiometry

Measured Response:

Scintillation

Testing Information

Procedure Summary:

This assay measures binding of [³H] Colchicine to rat colchicine binding protein. Whole brain membranes of Wistar derived rats weighing 175 ± 25 g are prepared in Na phosphate pH 6.7 buffer using standard techniques. A 0.1 mg‡ aliquot of membrane is incubated with 2.5 nM [³H] Colchicine for 3 hours at 37°C. Non-specific binding is estimated in the presence of 100 μM Colchicine. Membranes are filtered and washed 3 times and the filters are counted to determine [³H] Colchicine specifically bound. Compounds are screened at 10 μM.
Note: ‡Membrane protein may change from lot to lot, the concentration used will be adjusted if necessary.

Ligand:

[³H] Colchicine

Ligand Kd (nM):

1200

Ligand Concentration:

2.5 nM

Non Specific:

100 µM Colchicine

Incubation:

180 min at 37°C

Control Inhibitor:

Colchicine

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

6167

Name

Colchicine

Measurement

2.8μM - IC50

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei