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Table 3 Tabular form of IHC

From: Deficient expression of DNA repair enzymes in early progression to sporadic colon cancer

Pms2 Ercc1 Xpf Ku86
Cut 4 micron sections Cut 4 micron sections Cut 4 micron sections Cut 4 micron sections
Deparaffinize and rehydrate Deparaffinize and rehydrate Deparaffinize and rehydrate Deparaffinize and rehydrate
Antigen retrieval in microwave with VECTOR Antigen Unmasking Solution Antigen retrieval in microwave with VECTOR Antigen Unmasking Solution Antigen retrieval in microwave with VECTOR Antigen Unmasking Solution Antigen retrieval in microwave with VECTOR Antigen Unmasking Solution
Rinse with PBS and distilled water Rinse with PBS and distilled H2O Rinse with PBS and distilled water Rinse with PBS and distilled water
Incubate in 3% H2O2 in methanol for 20 min Incubate in 3% H2O2 in methanol for 20 min Incubate in 3% H2O2 in methanol for 20 min Incubate in 3% H2O2 in methanol for 20 min
Rinse in distilled water, PBS Rinse in distilled water, PBS Rinse in distilled water, TBST, PBS Rinse in distilled water, PBS
Place in Sequenza racks, rinse with PBS Place in Sequenza racks, rinse with PBS Place in Sequenza racks, rinse with PBS Place in Sequenza racks, rinse with PBS
Add 3 drops Background Sniper, 10 min at room temp Add 3 drops Background Sniper, 10 min at room temp Add 3 drops Background Sniper, 10 min at room temp Add 1.5% normal goat serum in 2%
BSA/PBS, 120 μl, one hour
Rinse with TBST Rinse with PBS Rinse with TBST Do Not Rinse
Primary mouse monoclonal from Pharmingen at 10 μg/ml into 2% BSA/TBST, 120 μl/slide Primary mouse monoclonal 8F1 from Neomarkers at 2 μg/ml with 2% BSA/PBS, 120 μl/slide Primary rabbit polyclonal 3F2/3 from Abcam at 5 μg/ml in Renoir Red at 120 μl/slide Primary mouse monoclonal H-300 from Santa Cruz at 2 μg/ml in 2% BSA/PBS, 120 μl/slide
Incubate 2 hrs at room temp Incubate 45 min at room temp Incubate 1 hour at room temp Incubate 45 min at room temp
3 rinses with TBST 3 rinses with PBS 3 rinses with TBST 3 rinses with PBS
Polyclonal rabbit anit-mouse antibody Dako Biotinylated secondary antibody at 100 μl/slide at 1:100 dilution in 2% BSA in TBST Polyclonal rabbit anit-mouse antibody Dako Biotinylated secondary antibody at 100 μl/slide at 1:300 dilution in 2% BSA/PBS Mouse Probe (Biocare) 4 drops/slide Goat anti-rabbit biotinylated IgG secondary antibody at 1:100 dilution in 2% BSA/PBS, 100 μl/slide
Incubate 30 min at room temp Incubate 30 min at room temp Incubate 15 min room temp Incubate 30 min at room temp
3 rinses with TBST 3 rinses with PBS 3 rinses with TBST 3 rinses with PBS
Vectastain Elite avidin-biotin complex method kit 3 drops/slide Vectastain Elite avidin-biotin complex method kit 3 drops/slide Mouse Polymer (Biocare)
4 drops/slide
Vectastain Elite avidin-biotin complex method kit 3 drops/slide
Incubate 30 min room temp Incubate 30 min room temp Incubate 15 min room temp Incubate 30 min room temp
2 rinses TBST, then PBS 2 rinses PBS 3 rinses TBST, then PBS 2 rinses PBS
Diaminobenzidine tetrachloride plus 0.04% H2O2 Diaminobenzidine tetrachloride plus 0.04% H2O2 Diaminobenzidine tetrachloride plus 0.04% H2O2 Diaminobenzidine tetrachloride plus 0.04% H2O2
Counterstain with hematoxylin Counterstain with hematoxylin Counterstain with hematoxylin Counterstain with hematoxylin
Dehydrate with ethanols followed by xylene Dehydrate with ethanols followed by xylene Dehydrate with ethanols followed by xylene Dehydrate with ethanols followed by xylene
Mount with Cytoseal and coverslips Mount w/Cytoseal and coverslips Mount with Cytoseal and coverslips Mount with Cytoseal and coverslips