Ismail Amr El-ShimyHumboldt-Universität zu Berlin | HU Berlin · Integrative Research Institute for the Life Sciences (IRI)
Ismail Amr El-Shimy
Bachelor degree in Pharmacy and Biotechnology, Master of Science degree in Molecular Pharmacology
About
7
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Introduction
Skills and Expertise
Publications
Publications (7)
As the death toll of Coronavirus disease 19 (COVID‐19) continues to rise worldwide, it is imperative to explore novel molecular mechanisms for targeting SARS‐CoV‐2. Rather than looking for drugs that directly interact with key viral proteins inhibiting its replication, an alternative and possibly add‐on approach is to dismantle the host cell machin...
Oncogenic mutations in KRAS or BRAF are frequent in colorectal cancer and activate the ERK kinase. Here, we find graded ERK phosphorylation correlating with cell differentiation in patient-derived colorectal cancer organoids with and without KRAS mutations. Using reporters, single cell transcriptomics and mass cytometry, we observe cell type-specif...
Mutations activating the KRAS GTPase or the BRAF kinase are frequent in colorectal cancer. Here, we use inducible transgenic expression of KRASG12V or BRAFV600E in intestinal organoids of mice to investigate oncogenic signal transduction in the mitogen-activated protein kinase (MAPK) cascade with cellular resolution. Using phospho-protein, reporter...
Introduction
The epidermal growth factor receptor (EGFR) signalling pathway has been established as a key driver of cell proliferation, growth and migration. Activating mutations as well as amplifications of the receptor gene have been found in a number of solid tumours such as glioblastoma, non-small cell lung cancer and colorectal cancer. For thi...
Microglia, the brain innate immune cells, are activated in response to amyloid beta (Aβ) resulting in neuroinflammation in AD brains. Recently, two phenotypes have been described for microglia: the pro-inflammatory classical and the anti-inflammatory alternative. Changes in microglia phenotype that control their phagocytic function are yet to be de...
Objectives
Neuroinflammation has been established as a fundamental component of Alzheimer’s disease (AD) pathogenesis. Microglia, the brain innate immune cells, are activated in response to amyloid beta (Aβ) leading to neuroinflammation in AD. Recently, two phenotypes have been described for microglia: proinflammatory classical M1 and anti-inflamma...
Questions
Questions (3)
I was looking for an antibody that can block the ligand binding domain of mouse EGFR. Can I use Cetuximab or is it exclusively human-specific?
I need a general protocol for the assay. How long should I incubate microglia with the tested compound?
I have tried incubating the cells for 24 hrs with the drug, but the problem I have is that the absorbance of the blank (DMEM culture medium without any cells) is always greater than the absorbance of the samples containing cells. This is very strange because the absorbance produced by cells should be significantly higher than that produced by DMEM medium alone.
I have seeded 50,000 cells per well in a 96 well plate and incubated these for 24 hrs with the drug. Then I removed the drug containing the medium and added fresh medium (100 ul per well) together with 10 ul of WST-1 reagent per well and incubated the cells for another 24 hrs. Then I measured the absorbance of the samples and to my surprise the absorbance of the blank (DMEM alone) was greater than the absorbance of all samples containing cells. Has anyone had a similar problem? Do you have any suggestions?
Hint: I use primary microglial cell cultures that are freshly isolated from mouse brain tissue
Microglia isolation using percoll
