To obtain the serum from a mouse blood sample I perform this protocol:
1. Obtain the sample withouth any anticoagulant and transfer to a strile empty tube.
2. Leave the tube in a standing position and wait 30 min.
3. Centrifuge 1500 g 10 min at 4ºC.
4. Take out the serum.
5. Keep the sample at 4ºC up to 6 months.
Has somebody a better protocol for this? Can I keep the sample at -20ºC or -80ºC thus I can store it more time?
Hello Laura
To get serum from mouse blood, we usually centrifuge our samples at low speed (we use 1000rpm for 5 min for 2mL blood sample but the rpm and time depend on the volume of blood). The idea is to spin at low rpm to avoid lysis of RBC (which will completely ruin your serum).
Serum is basically rich in proteins, hence if you wish to use is in the future for protein work, it is highly discouraged to freeze it, since freezing-thawing will disturb the proteins in the serum and the overall sample quality.
If the serum sample is ultra precious and you desperately need super long time storage, try to lyophilize/freeze-dry your sample. This process is also used to make commercial BSA. Nonetheless, there is no guarantee that all protein integrity will be 100% preserved. So far 4deg C is the least harmful though short lived.
Hope this helps
I recommend, before freezing, add protease inhibitor cocktail to the sample.
Hello Dr. Laura Díaz Marugán ,
I think, after collecting blood from mouse without anticoagulant, you should keep it for few hours and then , start centrifuging from 3000 rpm to 10000 rpm and each time, keep collecting cell free supernatant.
Regards,
Babu
- Badges
- Science topic
- Similar topics
- Medicine
- Disease
- Disease Models
- Animal Models
- Rodent Models
- Mouse Models