Journal
APPLIED IMMUNOHISTOCHEMISTRY & MOLECULAR MORPHOLOGY
Volume 17, Issue 2, Pages 165-171Publisher
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1097/PAI.0b013e3181853001
Keywords
protein snap-frozen heat-dried; desiccated; protein extraction; molecular biology; immunohistochemistry
Funding
- NCCAM [5R24AT002681]
- NEI [5P30EY603040]
- Arnold and Mabel Beckman Foundation
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Cryosectioned tissues from snap-frozen samples offer the advantage of preserving proteins at the cellular and subcellular levels and maintaining overall cell integrity in the tissue of interest without the use of chemical fixatives. To prevent specific or nonspecific degradation of proteins by autolytic and/or proteolytic processes, it is common practice to immediately store frozen tissue sections obtained from a cryostat under cryogenic conditions, for example -80 degrees C. Our laboratory recently challenged this widely held belief by extracting proteins from brain tissue samples that were archived for 1 day, 1 week, 1 month, and 6 months at various storage conditions (frozen, ambient, or desiccated) without the use of chemical fixatives. Our results from immunofluorescent stains, immunoperoxidase stains, silver stains, and Western blot analyses demonstrated that snap-frozen, heat-dried tissue sections stored and desiccated at ambient laboratory conditions are comparable to frozen samples stored up to 6 months.
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