Protein profiling in cancer cell lines and tumor tissue using reverse phase protein arrays
نویسندگان
چکیده
Reverse phase protein array (RPPA) technology is an antibody-based high-throughput assay for protein profiling of biological specimens that allows for many measurements with very small amounts of cell lysate. Here, we report the sensitivity, reproducibility, and accuracy of a particular RPPA platform called Zeptosens. We customized the RPPA protocol for our in-house setup, and measured more than 80 total protein and phospho-protein levels in various cancer samples, including cell lines, organoids, tumor chunks, core needle biopsies, and laser-capture microdissected tissue samples. We discuss pros and cons of the RPPA platform, and describe results from profiling 15 cancer cell line cells using RPPA.
منابع مشابه
Proteomic profiling of the NCI-60 cancer cell lines using new high-density reverse-phase lysate microarrays.
Because most potential molecular markers and targets are proteins, proteomic profiling is expected to yield more direct answers to functional and pharmacological questions than does transcriptional profiling. To aid in such studies, we have developed a protocol for making reverse-phase protein lysate microarrays with larger numbers of spots than previously feasible. Our first application of the...
متن کاملExpression Status of UBE2Q2 in Colorectal Primary Tumors and Cell Lines
Background: Activation of the ubiquitin-proteasome pathway in various malignancies, including colorectal cancer, is established. This pathway mediates the degradation of damaged proteins and regulates growth and stress response. The novel human gene, UBE2Q2, with a putative ubiquitin-conjugating enzyme activity, is reported to be overexpressed in some malignancies. We sought to investigate the ...
متن کاملProteomic analysis of breast cancer molecular subtypes and biomarkers of response to targeted kinase inhibitors using reverse-phase protein microarrays.
Although breast cancer molecular subtypes have been extensively defined by means of gene expression profiling over the past decade, little is known, at the proteomic level, as to how signaling pathways are differentially activated and serve to control proliferation in different breast cancer subtypes. We used reverse-phase protein arrays to examine phosphorylation status of 100 proteins in a pa...
متن کاملبررسی میزان بیان تلومراز در دو رده سلولی آلوده به ویروس پاپلیومای انسانی پرخطر
Background: Human papilloma virus (HPV) is one of the most important factors in cervical cancer. Viral sequences are integrated into the host cell genome. In mild cases the virus causes skin damages, in severe cases it leads to cancer. Like many other cancers, telomerase gene expression was increased in cervical cancer. This enzyme is a reverse transcriptase that contains two common subunits: i...
متن کاملDown-regulation of HSP40 gene family following OCT4B1 suppression in human tumor cell lines
Objective(s): The OCT4B1, as one of OCT4 variants, is expressed in cancer cell lines and tissues more than other variants and plays an important role in apoptosis and stress (heat shock protein) pathways. The present study was designed to determine the effects of OCT4B1 silencing on expressional profile of HSP40 gene family expression in three different human tumor cell lines. Materials and Met...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره شماره
صفحات -
تاریخ انتشار 2017