I need to store fresh frozen tumor tissue from nude mice for western blotting, receptor binding assays, and immunohistochemistry. In our lab, we freeze tissue for western blots in RIPA buffer, in TRIS/all for RBAs, and flash freeze in 2methylbutane + dry ice for IHC. I will not be able to analyze the tissue for some time and was wondering how long it will stay good at -80C in that manner.
According to AATB, the expiration time of different human tissue should not exceed the following:
• Refrigerated musculoskeletal human tissue: 5 days;
• Refrigerated human skin: 14 days;
• Frozen and cryopreserved cells and human tissue (-40 C or colder): 5 years;
• Lyophilised or dehydrated human tissue: 5 years.
Safe duration of storage of autographs may be influenced by processing and packaging methods and should be defined by the facility, not to exceed the expiration date listed above. A process should be in place to minimize the number of autographs in storage to those that will be reimplanted. This is facilitated by periodic communication with surgeons or review of medical records to determine whether patients have survived.
It is important to be sure that the storage conditions are the appropriate to maintain the quality of the processed tissue. However, you should check this information taking into account the specific type of tissue you that you intend to storage.
According to AATB, the expiration time of different human tissue should not exceed the following:
• Refrigerated musculoskeletal human tissue: 5 days;
• Refrigerated human skin: 14 days;
• Frozen and cryopreserved cells and human tissue (-40 C or colder): 5 years;
• Lyophilised or dehydrated human tissue: 5 years.
Safe duration of storage of autographs may be influenced by processing and packaging methods and should be defined by the facility, not to exceed the expiration date listed above. A process should be in place to minimize the number of autographs in storage to those that will be reimplanted. This is facilitated by periodic communication with surgeons or review of medical records to determine whether patients have survived.
It is important to be sure that the storage conditions are the appropriate to maintain the quality of the processed tissue. However, you should check this information taking into account the specific type of tissue you that you intend to storage.
In all labs I have worked in there was tissue storage (both snap-frozen biopsies as RIpA lysed cells for western blot) at -80 for substatial time: 5 moths up to one year, and it seemed to work fine. Specially iff you allready lysed the tissue for western blots, I think you can safely put the samples in -80 for quite some time.
Hi Beth
I agree with Jorge. In my experience, flash frozen tissue works quite fine in western blots even if stored for extended periods like 2-3 years at -80 degree. However, for IHC the flash frozen technique is not good (we freeze in liq. N2). We fix the freshly captured sample in 4% PFA, process it with sucrose gradient and finally embed in OCT compound. This form stays good for 2-3 yrs at -80 degree.
Hope it helps!
Agree with the above suggestions but it will be hard to store the tissue for receptor binding assays and immunohistochemistry. For IHC you need to have a small piece stored in PFA or formalin. I have no experience with 2-methylbutane for IHC. However, it will be important to find out how you want to study the receptor binding. Different storage method will be needed for specific way to study it.
Hi!!
In our lab we keep the frozen tissue samples in -80 deep freezer for long time such as 2-3 years..You can freeze your tissue sample in liquid nitrogen as well..but you have to take few precaution during the thawing. so try any of these two methods and your sample will be safe for a long time..
Thank you all for your responses! I only have to store it for about a month or two, so this makes me feel much better about results. I will be embedding the flash frozen tissue in OCT as well. Thank you.
Be careful that freeze-thaw process could make some changes-both protein levels and post-translational modifications. We have met such kind of problems in our lab. So my suggestion is for samples which will be subjected to western blot, you'd better lyse them immediately after taking from the nude mice, instead of store them at -80 directly. It is the best way to keep the samples as their original physiological/pathological states. The lysate could stay up to 6 months without detectable changes.
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