The formalin-fixed paraffin-embedded tissue sections were treated with xylene, graded alcohol and then antigen retrieval was performed in 0.01 M citrate buffer. Hydrogen peroxide was used for blockage. Tissue sections were treated with goat serum for 20 min. Sigma Prestige SIX1 antibody (1:300; Sigma-Aldrich Co., St Louis, MO, USA) was incubated with each section at 4°C overnight. EliVision plus and DAB kits from Maixin (Fuzhou, China) were used for immunohistochemical staining. Staining was performed according to manufacturer’s procedure. All tumor slides were examined randomly by two independent pathologists. Staining in the nucleus was considered as SIX1 positive staining. Staining intensity was classified as: 0: no staining, 1: moderate staining, and 2: strong staining and staining percentage was scored as: 1: 1%–24%, 2: 25%–49%, 3: 50%–74%, and 4: 75%–100%. Percentage score and intensity score were multiplied to give a final SIX1 score. Tumors with a final score ≥4 were considered as SIX1 overexpressed tumors.
Elivision plus
Manufactured by Maixin Group
Sourced in China, United States
EliVision plus is a high-performance laboratory instrument designed for automated analysis. It is capable of conducting a variety of analytical tasks with precision and efficiency.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using elivision plus
1
Immunohistochemical Analysis of SIX1 Expression
2
Immunohistochemical Staining Protocol for Tumor Samples
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The immunohistochemistry detection kit (EliVision plus) and DAB staining kit were purchased from Maixin Biotechnology Company in Fuzhou, China. Formalin was utilized to preserve all the tumor samples. To prepare the tissues for staining, they were first sectioned to a thickness of 5 micrometers and then put on glass slides, followed by routine dehydration, paraffin embedding, and consecutive sectioning with a thickness of 4μm. Deparaffinization was done using xylene, followed by gradient ethanol hydration. EDTA high-temperature high-pressure antigen retrieval, DAB staining, and counterstaining with hematoxylin were performed. The primary antibody was diluted at a concentration of 1:1000. Immunohistochemical staining was performed using the EnVision two-step method, and all experimental procedures strictly followed the instructions provided with the kit.
3
Quantifying Tumor Angiogenesis via CD34 IHC
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Sections cut from paraffin-embedded nude mice tumor samples were deparaffinized and rehydrated. Immunohistochemical staining assay was used to detected the expression of CD34 in all tumor sections with CD34 antibody (1:250, Abcam, USA) and Elivision™ plus (Maixin Biotechnology Co. Ltd, China). Then cell nucleus was stained with hematoxylin (Maixin Biotechnology Co. Ltd, China). After hydration and transparent, sections were sealed with neutral resins and photographed. Finally, CD34 positive vessels which were defined as blood vessels were analyzed with Image Pro Plus.
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