TGF-f3/Smad signaling pathway-related protein expression
TGF-f3/Smad signaling pathway-related protein expression
Transforming growth factor (TGF)-13/Smads signaling pathway related protein expression is mainly used to: (1) study the expression of related proteins in the TGF-13/Smad signaling pathway in tumor tissues; (2) elucidate the pathogenesis of cancer by correlating with clinicopathological data for relevant analysis.
Operation method
TGF-f3/Smad signaling pathway-related protein expression
Principle
Abnormalities in any component of the TGF-13/Smads signaling pathway can cause TGF-13/Smads signaling disorders, which can lead to tumorigenesis. Studying the expression of related proteins in the TGF-13/Smads signaling pathway in tumor tissues and correlating them with clinicopathological data for relevant analysis can elucidate the pathogenesis of cancer.
Materials and Instruments
Autopsy Pancreas Specimen Move I. Collection of case data Surgically resected and autopsied pancreatic specimens were collected from January 2001 to December 2003 in the Department of Pathology, Changhai Hospital, Second Military Medical University, with more complete data, totaling 250 cases. All the pathologic sections were reviewed, and according to the latest concept and diagnostic criteria of PanlN, all the PanlN foci in the sections were detected, totaling 351 foci, distributed in 156 cases. These 156 cases included 121 cases of ductal adenocarcinoma, 23 cases of chronic pancreatitis, and 12 cases of normal pancreas. There were 93 males and 63 females, aged 3O--8O years, with a mean of 58.6 years.Of the 121 cases of pancreatic ductal adenocarcinoma, 92 cases occurred in the head of the pancreas, 27 cases in the tail of the pancreas, and 2 cases in the whole pancreas. Pathologically, 21 cases were highly differentiated, 68 cases were moderately differentiated, and 32 cases were poorly differentiated. II. Tissue microarray construction We identified the HE sections with PanlN foci and the corresponding archived tissue wax blocks, marked the PanlN foci on the sections with a marker, and then compared the wax blocks and marked the corresponding points. Tissue microarrays were prepared using a tissue microarrayer (Tissue Arrayer, Beecher Instruments, Silver Spring, MD, USA) according to the method established by Kononen et al. The procedure was as follows: 1. The constructor was used to perforate holes (2 mm in diameter and 2--3 mm in depth) in a pre--prepared empty white wax block (45 mm x 20 mm), and then perforate the tissue at the marked points of the tissue wax block (the same diameter and depth as above), and accurately put it into the holes of the empty block, and then operate in a sequential order until the completion of the process, and then record the results. 2. The prepared chip wax block is then processed so that the embedded tissue strips are closely fused with the wax block. 3. Finally, a paraffin slicer was applied for continuous sectioning. Immunohistochemical staining 2--4 m thick paraffin sections were used for immunohistochemistry by S-P method and EnVision two-step method, with PBS replacing the primary antibody as negative control, and DAB developing the color.Smad2,3,4,7 antibodies were purchased from Santa Cruz, TGF.131, TGF.13R U antibodies were purchased from Wuhan Bode company, TGF-13R U antibody was purchased from Wuhan Bode company, and TGF-13R U antibody was purchased from Wuhan Bode company. TGF.131 and TGF.13R U antibodies were purchased from Wuhan PhD Company, and TGF-13R I antibody was purchased from Cell Signaling Technology Company. Determination of results The results of the staining were based on the criteria in the relevant literature with slight modification. Smad2, 3 and TGF-131 were positive for the presence of obvious brownish-yellow particles in the cytoplasm; Smad4 and 7 were positive for the presence of obvious brownish-yellow particles in the cytoplasm and/or the nucleus; and TGF-13R I, and TGF-13RII were positive for the presence of brownish-yellow coloration in the cytosol and/or the cytoplasm. Positive cases were recognized if the number of positive cells was >10%, and negative cases if the number of positive cells was ≤10%. V. Statistical processing The data were analyzed by x2 test using SPSS statistical software. Common Problems The steps of this experiment refer to the content of "Systematic Comparison of Related Protein Expression in the TGF-13/Smad Signaling Pathway in PanIN and Pancreatic Cancer Tissues of Different Grades by Tissue Microarray and Immunohistochemistry Techniques". For more product details, please visit Aladdin Scientific website.
Smad Antibody TGF-13R I Antibody
Paraffin Sections Tissue Microarray Constructor