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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