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Citric acid antigen retrieval (CAAR) for tryptic peptide imaging directly on archived formalin-fixed paraffin-embedded tissue

, : Citric acid antigen retrieval (CAAR) for tryptic peptide imaging directly on archived formalin-fixed paraffin-embedded tissue. Journal of Proteome Research 9(9): 4315-4328

Imaging mass spectrometry (IMS) is a powerful technology for mapping distributions of biological molecules like proteins and peptides within tissue sections. It is therefore potentially extremely useful for the analysis of pathological conditions such as neoplastic diseases. The use of IMS is typically limited to fresh frozen tissue specimens. However, there is a high interest in the possibility of being able to analyze the tissue proteome of formalin-fixed paraffin-embedded (FFPE) specimens that have been stored together with the clinicopathological information of patients in huge archives over many decades. We have therefore developed an antigen-retrieval protocol using a high temperature citric acid buffer to allow partial reversal of FFPE protein cross-linking. Coupled with automated deposition of trypsin and matrix, our method allows the generation of meaningful peptide ion distribution images. In situ peptide fragmentation provided identification of high abundance proteins such as Actin and Collagen. Furthermore, downstream application of three different HPLC-MS strategies allowed identification of a maximum of 106 proteins, 67 of which were mass correlated to ions from IMS analysis of archived FFPE ovarian tissue. The CAAR method presented here complements previously described antigen-retrieval protocols and is an important step in being able to fully analyze the proteome of archived FFPE tissue.


PMID: 20597554

DOI: 10.1021/pr9011766

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Koshy, P.J.; Rowan, A.D.; Life, P.F.; Cawston, T.E., 1999: 96-Well plate assays for measuring collagenase activity using (3)H-acetylated collagen. We describe two alternative assays for measuring collagenolytic activity using (3)H-acetylated collagen. Both assays have been developed for the 96-well plate format and measure the amount of radiolabeled collagen fragments released into the super...

Kato, H.; Yamane, S.; Phengklai, C., 2004: Ant-colonized domatia on fruits of Mucuna interrupta (Leguminosae). Domatia of myrmecophytes have been reported only in vegetative organs so far. In this paper, we report the first record of domatia formed on the legume surface of Mucuna interrupta. The domatium consists of 12-16 vertical lamellae which traverse t...

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Mullen, G.R., 1977: Acarine parasites of mosquitoes. IV. Taxonomy, life history and behavior of Thyas barbigera and Thyasides sphagnorum (Hydrachnellae: Thyasidae). In this part of a series on mite parasites of mosquitoes [see RAE/B 63, 488, 3358], characters are provided for distinguishing the larvae of Thyas barbigera Viets and Thyasides sphagnorum Habeeb [see 63, 3358], which commonly parasitise mosquitoes...

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Mello, S., 1969: Assis Chateaubriand. Revista Brasileira de Medicina 26(1): 66-67

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Mastiukova, E.M., 1971: Study of internal speech in school children with different speech disorders. Zhurnal Nevropatologii i Psikhiatrii Imeni S.S. Korsakova 71(10): 1512-1517

de Bruin, A.; Treccani, B.; Della Sala, S., 2015: The connection is in the data: we should consider them all. Psychological Science 26(6): 947-949

Tabuenca, M.C., 1985: Reduction in fruit cracking in cherries following alpha -naphthylacetic acid (NAA) treatment. NAA application at 1 p.p.m. 25-30 days before harvest reduced the cracking index in cultivars Bing, Marmotte, Mollar de Caceres and Napoleon by 25-55%, but had no effect on fruit cracking in cultivars Daiber and Van.