Biomarker for breast cancer metastasis and prognosis diagnosis
A biomarker, breast cancer technology, applied in the field of biomedicine
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0049] Example 1 Preparation of antibodies for detecting meR342-EZH2
[0050] The antibody used to detect meR342-EZH2, the amino acid sequence of the antibody is shown in SEQ ID No.1 (TAERIKTPPKRPGGC).
[0051] A method for preparing an antibody for detecting meR342-EZH2, comprising the steps of:
[0052] (1) Using polypeptide synthesis technology to synthesize a polypeptide with an amino acid sequence such as the antibody shown in SEQ ID No.1, so that the purity of the polypeptide is above 85%, and then cross-linking with hemocyanin KLH to obtain the antigen;
[0053] (2) The antigen is mixed with complete adjuvant (CFA) or incomplete adjuvant (IFA) in equal volumes, injected into the animal body, immunized 8 times, and the whole blood of the animal is taken for purification to obtain the antibody.
[0054] In step (2), the animals are New Zealand white rabbits. Take two adult male New Zealand white rabbits purchased from Weitong Lihua Company (the numbers are RB11081 and RB...
Embodiment 2
[0064] Example 2 The relationship between the expression of meR342-EZH2 and the metastasis of breast cancer
[0065] Get 100 cases of breast cancer tissues (wherein, 10 cases of paracancerous tissues, 50 cases of breast cancer tissues in situ, 40 cases of lymph node metastasis breast cancer tissues), adopt the antibody prepared in Example 1, and pass immunohistochemical experiments (using Zhongshan Jinqiao The IHC kit was completed (Zhongshan Jinqiao PV600)) to detect the expression of meR342-EZH2 in normal breast tissue, primary breast cancer samples and metastatic breast cancer samples.
[0066] The operation of the immunohistochemical experiment is as follows:
[0067] (1) Conventional paraffin sections, dewaxed to water, 3% H 2 o 2 Incubate at room temperature for 10 minutes to eliminate endogenous peroxidase activity, wash with PBS 3 times, 5 minutes / time, then boil with antigen retrieval solution (EDTA / sodium citrate) in a pressure cooker for 5 minutes, naturally cool ...
Embodiment 3
[0077] Example 3 meR342-EZH2 promotes the EMT process of breast cancer cells and the invasion and migration of breast cancer cells
[0078] (1) meR342-EZH2 promotes the EMT process of breast cancer cells
[0079] 1) Lentivirus packaging: Spread 293T cells into a 6cm cell culture plate the day before, so that the cell density before transfection reaches 80%; the ratio of related plasmids and transfection reagents for lentivirus packaging is the core plasmid (shEZH2-3'UTR / Vector / Flag-EZH2-WT / Flag-EZH2-R342K): PAX:PMD2G:PEI=4:3:1:3, the total amount of virus plasmid transfected on a 6cm plate is 8μg; add DMEM serum-free medium to a dorf tube After adding the plasmid and PEI, blow and mix gently, and let stand at room temperature for 15 minutes; 6 hours after transfection, discard the medium and replace it with 3ml DMEM complete medium. Collect the first batch of virus liquid after 48 hours of transfection, and then every 24 hours Collect once, replace with 3ml of fresh DMEM com...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com