Please note which sample types are recommended in the manual for the kit you are interested in. If you have any questions or would like to know if we can make a custom kit to fit your needs, please contact us.
Use a serum separator tube and allow samples to clot for two hours at room temperature or overnight at 4℃ before centrifugation for 20 minutes at approximately 1000×g. Assay freshly prepared serum immediately or store samples in aliquots at -20℃ or -80℃ for later use. Avoid repeated freeze/thaw cycles.
Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2 - 8℃ within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquots at -20℃ or -80℃ for later use. Avoid repeated freeze/thaw cycles.
The preparation of tissue homogenates will vary depending on tissue type. In general, tissues should be rinsed thoroughly in ice-cold PBS (0.02mol/L,pH 7.0-7.2) to remove excess blood and weighed before homogenization. Mince the tissues to small pieces and homogenize them in 5-10 mL of PBS with a glass homogenizer on ice (Micro Tissue Grinders work as well). The resulting suspension should be sonicated with an ultrasonic cell disrupter or subjected to two freeze-thaw cycles to further break the cell membranes. After that, the homogenates should be centrifugated for 5 minutes at 5000×g. Remove the supernatant and assay immediately or aliquot and store at ≤-20 ℃.
Cells must be lysed before assaying according to the following directions:
- Adherent cells should be detached with trypsin and then collected by centrifugation (suspension cells can be collected by centrifugation directly).
- Wash cells three times in cold PBS.
- Resuspend cells in 1X PBS and subject to ultrasonication 4 times (or Freeze cells at≤-20℃. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle 3 times.)
- Centrifuge at 1500×g for 10 minutes at 2 - 8℃ to remove cellular debris. Cell culture supernates and Other biological fluids - Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediately or store samples in aliquot at -20℃ or -80℃. Avoid repeated freeze/thaw cycles.
Collect saliva using a collection device or equivalent and transfer to a centrifuge tube. Freeze sample at -70°C for 1 hour. Thaw on ice, and centrifuge at 2,000 × g for 10 minutes. Transfer clarified supernatant to clean tube for use. Assay immediately or store samples in aliquots at ≤-20oC. Avoid repeated freeze/thaw cycles.
Aseptically collect the first urine of the day (mid-stream), collect directly into a sterile container. Centrifuge to remove particulate matter, assay immediately or aliquot and store at ≤-20oC. Avoid repeated freeze-thaw cycles.
Centrifuge samples for 15 minutes at 10,000×g at 2 - 8oC. Collect the aqueous fraction and centrifuge twice more for a total of 3 cycles. Assay immediately or or store samples in aliquot at -20oC or -80oC for later use. Avoid repeated freeze/thaw cycles.
Tears, Cell culture Supernatant and Other Biological Fluids
Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediately or store samples in aliquots at -20oC or -80oC. Avoid repeated freeze/thaw cycles.
- Samples to be used within 5 days may be stored at 4oC, otherwise samples must be stored at -20oC (≤1 month) or -80oC (≤2 months) to avoid loss of bioactivity and/or contamination.
- Sample hemolysis will influence the results; hemolytic specimen used as samples will not be detected.
- When performing the assay, bring samples to room temperature.