Journal of Pathology and Translational Medicine

Original Articles

Evaluation of Protein Expression in Housekeeping Genes across Multiple Tissues in Rats: Hye Jeong Kim, Jong In Na, Byung Woo Min, Joo Young Na, Kyung Hwa Lee, Jae Hyuk Lee, Young Jik Lee, Hyung Seok Kim, Jong Tae Park; Korean J Pathol. 2014;48(3):193-200. Published online June 26, 2014; DOI: https://doi.org/10.4132/KoreanJPathol.2014.48.3.193

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Background

Housekeeping genes, which show constant protein expression patterns between different tissue types, are very important in molecular biological studies as an internal control for protein research.

Methods

The protein expression profiles of seven housekeeping genes (HPRT1, PPIA, GYS1, TBP, YWHAZ, GAPDH and ACTB) in various rat tissues (cerebrum, cerebellum, cardiac ventricle and atrium, psoas muscle, femoral muscle, liver, spleen, kidney, and aorta) were analyzed by Western blot and compared by coefficient of variation (CV).

Results

HPRT1 was stably expressed (CV≤10%) in six tissues (cerebrum, cerebellum, ventricle, femoral muscle, spleen, and kidney), PPIA was stably expressed in five tissues (cerebrum, cerebellum, ventricle, spleen and kidney), YWHAZ was stably expressed in three tissues (cerebrum, cerebellum, and kidney), and GAPDH was stably expressed in four tissues (cerebrum, ventricle, psoas muscle, and kidney). In comparison, GYS1, TBP, and ACTB were found to have CV values over 10% in all tissues. Of the seven genes examined, four (HPRT1, PPIA, YWHAZ, and GAPDH) were found to be stably expressed across multiple organs, with low CV values (≤10%).

Conclusions

These results will provide fundamental information regarding internal controls for protein expression studies and can be used for analysis of postmortem protein degradation patterns in forensic medicine.

Citations

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Quality Control Program for Fresh Frozen Tissue and Its Results of Chonbuk National University Hospital National Biobank of Korea.: Shin Young Park, Hyun Ah Baek, Hyoung Jong Kwak, Sang Hyun Hong, Ho Sung Park, Kyu Yun Jang, Woo Sung Moon, Myoung Jae Kang, Dong Geun Lee, Myoung Ja Chung; Korean J Pathol. 2010;44(3):295-301.; DOI: https://doi.org/10.4132/KoreanJPathol.2010.44.3.295

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Abstract PDF

BACKGROUND
Molecular tools for tissue profiling generally require collection of fresh frozen tissues (FFT) as sources of high-quality DNA and RNA. Nowadays, researchers carry out large-scale, multi-center studies and they request inter-institutional minimal intrinsic bias, some fundamental similarities, and the same standardized and validated procedures.
METHODS
This study reports standardized quality control procedure for fresh frozen tissue of the National Biobank of Korea.
RESULTS
The main procedures for quality control for FFT are as follows: records related to sample collection such as labeling of samples, transport temperature, lag time from excision of tissue to freezing, and sample size were reviewed for all fresh frozen samples. The stability of RNA and DNA in fresh frozen tissue was evaluated for 3% of collected samples and purity was assessed (ratio of the absorbance at 260 and 280 nm) as was integrity (agarose gel electrophoresis). Stained hematoxylin and eosin sections were reviewed by a pathologist to confirm the diagnosis and to assess how representative the frozen sample was.
CONCLUSIONS
We introduced that the quality-control criteria for fresh frozen tissue of the NBK. We expect that this study contributes to standardization of collection, storage, and quality control of fresh frozen tissue.

Citations

Citations to this article as recorded by

Influence of Cold Ischemia Time and Storage Period on DNA Quality and Biomarker Research in Biobanked Colorectal Cancer Tissues
Min Gyoung Pak, Mee Sook Roh
Kosin Medical Journal.2020; 35(1): 26. CrossRef

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