Antibody composition for detecting pancreatic duct adenocarcinoma immunohistochemical marker protein compositions and application of antibody composition
A technology of pancreatic ductal adenocarcinoma and antibody composition, which is applied in the directions of immunoglobulin, anti-peptide protease inhibitor immunoglobulin, measuring device, etc. There are problems such as reports of sensitive markers of lesions, which can reduce the difficulty of interpretation, overcome low timeliness and low specificity, and simplify the operation process.
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Embodiment 1
[0033] Cell culture and immunofluorescence staining: PANC-1 cell line (purchased from ATCC, USA) was grown in DMEM medium (purchased from GIBCO) containing 10% FBS. Cells in the logarithmic phase were inoculated in a 6-well plate, and sterilized coverslips were pre-laid in the well plate, 2×10 5 / hole. After the cells grew and adhered to the wall, the fixative solution was prepared with methanol / acetone at a ratio of 1:1 for cell fixation. Wash 2-3 times with PBS and block for 1 hour at room temperature in PBS containing 5% goat serum. The primary antibody was incubated overnight at 4°C. Wash with PBS 3 times, 5 minutes each time. Fluorescently labeled goat anti-mouse or goat anti-rabbit secondary antibody was incubated at room temperature for 2 hours. Wash with PBS 3 times, 5 minutes each time. Add anti-fluorescence quenching reagent, observe and record with laser confocal microscope; see results in figure 1 .
[0034] Paraffin section specimens: 2 cases of pancreatic ...
Embodiment 2
[0038]Paraffin section specimens: from 32 patients with pancreatic diseases hospitalized in the First Affiliated Hospital of Nanjing Medical University between January 2014 and January 2015, all of which were confirmed by pathological diagnosis. The median age was 65 years, including 18 male patients and 14 female patients. The clinical data are shown in Table 1. Among them, there were 28 cases of pancreatic ductal adenocarcinoma, 1 case of mucinous cystic tumor, 1 case of neuroendocrine tumor, and 1 case of serous tumor Cystadenoma, 1 case of mucinous adenocarcinoma. Of the 28 patients with pancreatic ductal adenocarcinoma, 14 had no lymph node metastasis and 14 had lymph node metastasis.
[0039] Table 1 Basic information of 32 patients with confirmed pancreatic diseases.
[0040]
[0041] Paraffin section immunohistochemistry: select 4 μm paraffin-embedded pancreatic tissue sections, select 4 μm paraffin-embedded pancreatic tissue sections, and incubate at 60° C. for 1 ...
Embodiment 3
[0044] Paraffin section specimens: from 10 patients with pancreatic diseases hospitalized in the First Affiliated Hospital of Nanjing Medical University between January 2014 and January 2015, all of which were confirmed by pathological diagnosis. The median age was 52 years old, including 5 male patients and 5 female patients. Among them, 5 cases of pancreatic ductal adenocarcinoma, 2 cases of solid-pseudopapillary tumor, 1 case of chronic pancreatitis, 1 case of pancreatic serous cystadenoma, and 1 case of neuroendocrine tumor. No lymph node metastasis occurred in 10 cases.
[0045] Paraffin section immunohistochemistry: select 4 μm paraffin-embedded pancreatic tissue sections, select 4 μm paraffin-embedded pancreatic tissue sections, and incubate at 60° C. for 1 hour. Soak in xylene for 30 minutes to dewax, and then soak in gradient ethanol solution in turn. Block with 3% hydrogen peroxide solution in methanol for 10 minutes at room temperature. Wash with PBS 3 times, 5 m...
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