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Real envision hrp rabbit mouse detection system

Manufactured by Agilent Technologies
Sourced in Denmark, United Kingdom

The Real Envision HRP Rabbit/Mouse detection system is a laboratory equipment product manufactured by Agilent Technologies. It is designed to detect the presence of rabbit or mouse proteins in biological samples using a horseradish peroxidase (HRP) enzyme-based detection method.

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2 protocols using real envision hrp rabbit mouse detection system

1

Immunohistochemical Analysis of Axin2 and Snail Expression in OSCC

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Immunohistochemistry was performed on 4-μm sections of paraffin-embedded tissue specimens of surgical margins obtained from 235 patients with OSCC. Axin2 (Rabbit polyclonal IgG, working dilution 1:500; ab32197, Abcam, Cambridge, UK) and Snail antibodies (Rabbit polyclonal IgG, working dilution 1:1000) were used as primary antibodies. The Snail antibody was prepared as previously described (25 (link)). Endogenous peroxidase activity was inhibited using a mixture of H2O2 and methanol (1:40), and antigen retrieval was performed by the pressure-cooking method using citrate buffer (pH 6.0, Sigma-Aldrich, Darmstadt, Germany) for the deparaffinized tissue sections. The Real Envision HRP Rabbit/Mouse detection system (Dako, Glostrup, Denmark) was used as a secondary antibody, and immunoreactivity of the tissue sections against Axin2 and Snail was visualized using 3,3′-diaminobenzidine.
The histoscore for the immunoreactivity of both Axin2 and Snail in the surgical margins of OSCC was calculated according to the staining intensity and percentage of positive cells using the weighted histoscore method (26 (link)). Patients were subdivided into two groups according to the histoscore: low (histoscore 0–100) and high (histoscore 101–300) expression groups.
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2

Immunohistochemistry for HIF-1α Expression

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The tissue samples were cut into 4-mm tissue sections for immunohistochemistry. The tissue sections were deparaffinized with xylene and hydrated using a graded ethanol. The mixture of H 2 O 2 and methanol (1:40 ratio) was used to inhibit endogenous peroxidase activity. Antigen retrieval was performed using an antigen retrieval solution (Dako, Carpinteria, CA), and a rabbit monoclonal IgG primary antibody to HIF-1a (Abcam, Cambridge, United Kingdom) (working dilution, 1:200) and a REAL EnVision HRP Rabbit/Mouse Detection System (Dako) secondary antibody were used in this study. Visualization was performed using the chromogen 3,3 0 -diaminobenzidine, and the counterstaining was performed using hematoxylin. Mouse IgG (DakoCytomation Denmark A/S, Glostrup, Denmark) was used as a negative control.
As described in our previous study, the weighted histoscore method was used to score HIF-1a expression according to staining intensity and the percentage of positively stained cells (Zhang et al., 2014) . The final score was calculated as follows:
final score¼ (0
The samples were subdivided into three groups, weak (histoscores 0e100), moderate (histoscores 101e200), and high HIF-1aeexpressing group (histoscores 201e300), based on the final histoscore.
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