PCNA Antibody (F-2)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Our specimens had been fixed with formalin from 24 to 48 h at room temperature and treated with routine processing as in a previous study (12). Ten 4-mm serial sections were prepared for staining and were incubated with primary antibodies for 2 h. Immunohistochemistry was performed using a Discovery Auto-Stainer with automated protocols (Ventana Medical Systems, Inc.; Roche) as previously described (13). The average number of PCNA-positive cells from case 1 to case 23 were counted in 10 random microscopic fields at 400 magnification.

Protocol tips

Our specimens had been fixed with formalin from 24 to 48 h at room temperature and treated with routine processing as in a previous study (12). Ten 4-mm serial sections were prepared for staining and were incubated with primary antibodies for 2 h. Immunohistochemistry was performed using a Discovery Auto-Stainer with automated protocols (Ventana Medical Systems, Inc.; Roche) as previously described (13). The average number of PCNA-positive cells from case 1 to case 23 were counted in 10 random microscopic fields at 400 magnification.

Publication protocol

Immunohistochemistry
Claudin-4, occludin, SOX2 and PCNA expression in BSCC and SCC tissues were evaluated from serial deparaffinized sections. Our specimens had been fixed with formalin from 24 to 48 h at room temperature and treated with routine processing as in a previous study (12). Ten 4-mm serial sections were prepared for staining and were incubated with primary antibodies for 2 h. Immunohistochemistry was performed using a Discovery Auto-Stainer with automated protocols (Ventana Medical Systems, Inc.; Roche) as previously described (13). The average number of PCNA-positive cells from case 1 to case 23 were counted in 10 random microscopic fields at 400 magnification. The SOX2 intensity was determined by qualitative assessment of three levels as weak, 1; moderate, 2; and strong, 3. We defined a diffuse staining pattern as that positively detected in almost all cancer cells but a dot-like staining pattern as that positively detected in partial and marginal cancer cells.
Western blot analysis
HSC2 cells were lysed using M-PER lysis buffer (Thermo Fisher Scientific, Inc.). Protein determination was performed using the bicinchoninic method. A total of 40 µg protein was loaded in each lane. The total cell lysates were run on 14% SDS-polyacrylamide gels followed by western blotting using standard procedures. The proteins were transferred on to PVDF membrane. For blocking, membranes were incubated with 5% skim milk for 60 min at room temperature after protein transfer. A WesternBright Sirius kit (Advansta) was used for antibody detection, and an AE-9300 Ez capture MG (ATTO) was used for image data capture. We repeated the western blot analysis three times and the results were similar.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Cell cytotoxicity / Proliferation assay cell type - oral squamous cell carcinoma using PCNA Antibody (F-2) from Santa Cruz Biotechnology.

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for PCNA Antibody (F-2) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Cell cytotoxicity / Proliferation assay cell type - oral squamous cell carcinoma using PCNA Antibody (F-2) from Santa Cruz Biotechnology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.