Isolation of high quality protein samples from punches of formalin fixed and paraffin embedded tissue blocks.
In general, it is believed that the extraction of proteins from formalin-fixed paraffin embedded samples is not feasible. However, recently a new technique was developed, presenting the extraction of non-degraded, full length proteins from formalin fixed tissues, usable for western blotting and protein arrays. In the study presented here, we applied this technique to punch biopsies of formalin fixed tissues embedded in paraffin to reduce heterogeneity of the tissue represented in sections, and to ensure analysing mainly defined cellular material. Successful extraction was achieved even from very small samples (0.7 mm(3)). Additionally, we were able to detect highly glycosylated proteins and protein modification, such as phosphorylation. Interestingly, with this technique it is feasible to extract high quality proteins from 14 year old samples. In summary, the new technique makes a great pool of material now usable for molecular analysis with high throughput tools.
Kroll
J
J
Institute of Pathology, University of Regensburg, Regensburg, Germany.
Becker
K F
KF
Kuphal
S
S
Hein
R
R
Hofstädter
F
F
Bosserhoff
A K
AK
D000951:Antigens, Neoplasm; D001706:Biopsy; D015153:Blotting, Western; D015820:Cadherins / Q000032:analysis / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005240:Feasibility Studies; D005404:Fixatives / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D005615:Freezing; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000601:surgery; D008545:Melanoma / Q000737:chemistry / Q000601:surgery; D008954:Models, Biological; D009363:Neoplasm Proteins; D016612:Paraffin Embedding; D011506:Proteins / Q000302:isolation & purification*; D016622:Silver Staining; D016707:Tissue Fixation; D025521:Tumor Suppressor Proteins / Q000032:analysis / Q000302:isolation & purification
D000951:Antigens, Neoplasm; D015820:Cadherins; D005404:Fixatives; C109278:MIA2 protein, human; D009363:Neoplasm Proteins; D011506:Proteins; D025521:Tumor Suppressor Proteins; D005557:Formaldehyde