Mice tongue tissues were fixed in 4% PFA, embedded in paraffin, and sectioned (4 μm) using standard procedures. IHC or IF staining was performed using an indirect method using primary antibodies recognizing YAP1 (WH0010413M1; Sigma), Ki67 (ab15580; Abcam), or TP63 (4A4; Abcam). Primary antibodies were detected using REAL EnVision HRP-rabbit/mouse (Dako) or Alexa Fluor 568 (Molecular Probes). Some slides were counterstained with Mayer’s hematoxylin (Muto) or 4′,6-diamidino-2-phenylindole (DAPI; Dojindo) before mounting using PermaFluor (Thermo Scientific). For Ki67 positivity studies, 200 cells per mouse were examined.
Real envision hrp rabbit mouse
Manufactured by Agilent Technologies
Sourced in United States
The REAL EnVision HRP-rabbit/mouse is a compact, automated slide staining platform designed for immunohistochemistry and in situ hybridization applications. It provides automated slide processing, including antigen retrieval, primary antibody incubation, and polymer-based detection. The instrument can accommodate both rabbit and mouse primary antibodies.
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Lab products found in correlation
3 protocols using real envision hrp rabbit mouse
1
Immunohistochemical Analysis of Tongue Tissues
2
Immunohistochemical and Histological Analysis
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Full-thickness skin samples from the experimental mice and human subjects were obtained and embedded in paraffin. The samples were cut into sections with a thickness of 5 mm and incubated with rabbit polyclonal anti-PAR-2 antibody (ab180953; Abcam) at a dilution ratio of 1:100 for 2 h. Then, staining with secondary antibody (Real™ Envision™ HRP Rabbit/Mouse; DAKO, Carpinteria, California, USA) was performed, and the sections were counterstained with hematoxylin.
For renal histology, kidney sections were stained with Masson's trichrome stain. The tubular injury score was scored semiquantitatively by an independent investigator who was blinded to the study design. Scoring was determined by assessing the cortex and corticomedullary junction as follows: 0, 0%; 1þ, <25%; 2þ, 25-50%; and 3þ, >50% of tubules. Randomly selected fields (n ¼ 10) from the cortex and corticomedullary junction in each kidney slide section were examined as described previously. 14
For renal histology, kidney sections were stained with Masson's trichrome stain. The tubular injury score was scored semiquantitatively by an independent investigator who was blinded to the study design. Scoring was determined by assessing the cortex and corticomedullary junction as follows: 0, 0%; 1þ, <25%; 2þ, 25-50%; and 3þ, >50% of tubules. Randomly selected fields (n ¼ 10) from the cortex and corticomedullary junction in each kidney slide section were examined as described previously. 14
3
Immunohistochemical Analysis of PTEN
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Formalin-fixed paraffin-embedded sections of tumor tissue were deparaffinized with xylene and hydrated with graded alcohol. Antigen retrieval was performed with a retrieval solution (DAKO, USA) using the pressure-cooking method, and the activity of endogenous peroxidase was blocked by a 1:40 mixture of hydrogen peroxide and methanol. The primary PTEN antibody (DAKO) incubation was performed at room temperature for 1 h in an antibody solution diluted to 1:100. All sections were incubated at room temperature for 30 min in the Real EnVision™ HRP Rabbit/Mouse (DAKO, USA) detection system, which functions as the secondary antibody. PTEN expression was indicated using a chromogen and counterstaining was performed with hematoxylin. PTEN expression was quantified by the H-score based on the intensity of cell staining and percentage of the stained cells [17 (link)]. Intensity was scored as 0: none, 1: weak, 2: moderate, or 3: strong. The H-score was calculated as follows: H-score = (%1 + cells × 1) + (%2 + cells × 2) + (%3 + cells × 3). An H-score of ≤ 10 was used as the cutoff point to define loss of PTEN expression based on a previous study.
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