Xenograft tumor tissues were dissected and fixed in 10% formalin, then were embedded and cut into 5 μm sections. The samples were deparaffinized, hydrated, stained with hematoxylin-eosin, then mounted using neutral resin. For immunohistochemical staining, endogenous peroxidase was quenched by 3% hydrogen peroxide and the sections were blocked with 2% goat serum at room temperature for 30 min, then incubated with antibodies for caspase-3, caspase-8, PARP-1, and CD31 (Abcam, UK) at 4°C overnight. The staining was detected using DBA kit (ZSGB-Bio, Beijing, China) and observed under a light microscope (Nikon DS-Ri1, Japan), the staining was quantified by Image J software.
Dba kit
Manufactured by ZSGB-BIO
Sourced in China
The DBA kit is a laboratory equipment designed for DNA binding assay. It provides the essential components required to perform DNA binding experiments in a research setting. The kit includes various reagents and solutions necessary for the assay, allowing researchers to analyze the interactions between DNA and target molecules.
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5 protocols using dba kit
1
Xenograft Tumor Histological Analysis
2
Immunohistochemical Analysis of Tumor Markers
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The tumor sections were dissected from mice, fixed in paraffin wax, and cut into sections of 5 μm thick. The sections were blocked with endogenous peroxidase activity with 3% hydrogen peroxide. After 30 min incubation with blocking serum, sections were incubated with antibodies for SPAG9 (1 : 200, Abcam, UK), E-cadherin (1 : 500, Abcam, UK), MMP-2 (1 : 500, Abcam, UK), and Vimentin (1 : 500, Abcam, UK) overnight at 4°C. The sections were stained by DBA kit (ZSGB-Bio, Beijing, China) and observed with light microscope (Nikon DS-Ri1, Japan). The sections were also counterstained with hematoxylin-eosin. ImageJ software was used to analyze the score of immunohistochemical staining which was expressed as mean density.
3
Apoptosis Protein Expression Analysis
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The sample in the paraffin was cut into a thickness of 4 μm. After deparaffinization and hydration, the sections were subjected to antigen retrieval. The endogenous peroxidase activity of the sections was blocked with 3% hydrogen peroxide. After incubation with blocking serum for 30 min, the sections were incubated with antibodies for Caspase-3 (1:500, Proteintech, USA), Caspase-8 (1:500, Proteintech, USA) and Bcl-2 (1:500, Abcam, Uk). After incubation at 4°C overnight, sections were stained with DBA kit (ZSGB-Bio, Beijing, China). The sections were observed under optical microscope (Nikon DS-Ri1, Japan) and analyzed by Image-J software.
4
Immunohistochemistry Analysis of Tumor Samples
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After mice were sacrificed for subcutaneous and intracranial tumour, we made specimens embedded with paraffin. Then the paraffin-embedded tumours were sectioned and dewaxed. After antigen retrieval using 10 mmol/l citrate buffer, sections were incubated with 3% H2O2 and blocked with 5% BSA. Then the sections were added with primary antibodies at 4 °C overnight. After rewarming at room temperature the next day, the sections were incubated with secondary antibodies using two-step polymer HRP detection system (ZSGB-BIO, Beijing, China). The samples were colourated with DBA Kit (ZSGB-BIO, Beijing, China) and then counterstained with haematoxylin. After dehydration and sealing piece with neutral gum, the samples were detected and photographed by microscope (Olympus Japan).
5
Immunohistochemical Analysis of Tumor Markers
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Tumor tissues were dissected from mice, fixed in 4% paraformaldehyde at 4°C overnight, and then embedded in paraffin wax, and finally cut into 5 μm thick sections. The sections were incubated with 3% hydrogen peroxide for 30 min at room temperature to block endogenous peroxidase activity. After 30 min incubation with blocking serum, sections were incubated with antibody for E-cadherin, vimentin or MMP9 (all from Abcam, Cambridge, UK; 1:500 dilution in phosphate-buffered saline pH 7.2 containing 0.1% bovine serum albumin) overnight at 4°C. The sections were stained with DBA kit (ZSGB-Bio, Beijing, China) and counterstained with hematoxylin-eosin. The stained sections were observed under microscope and the intensity of staining was analyzed by using Image-J software. Staining intensity was scored according to a prespecified semiquantitative intensity scale: −, negative with no staining; +, weak staining intensity in >10% tumor cells; ++, moderate staining intensity in >10% tumor cells; +++, strong staining intensity in >10% tumor cells
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