...Mitogen-activated protein kinases (MAPKs), also known as extracellular signal-regulated kinases (ERKs), are serine/...threonine-specific protein kinases. They regulate diverse cellular programs by acting as an integration point for...

...plasmids (AsCpf1 and LbCpf1) for use in mammalian cells SpCas9 variants with no detectable off-target effects...CRISPR-Cas/RGN expression plasmids for use in human cells CRISPR-Cas/RGN expression plasmids for use in zebrafish...

... tools that effectively transduce many mammalian cell types. AAV is used in a wide range of experiments...Tetracycline Transactivators, Affinity Purification, Cell Ablation Monosynaptic Neuronal Tracing Rabies Virus-Based...

...sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism...PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair ...

...sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism...PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair ...

...Fluorescent Protein Resources: Empty Backbones FRET Subcellular localization Optogenetics Background In vivo ...stable imaging of protein interactions in living cells and live mice. Proteins for in vivo imaging have...

...allowing a scientist to control signaling within cells expressing these engineered receptors. Chemogenetic...engineered receptor is coupled to and the specific cell type where the receptor is expressed, these receptors...

...Can I directly transfect cloning grade DNA into cells? Cloning grade DNA is designed for use in cloning...cloning applications. Direct transfection into cells is not recommended. Can I order more than 2 µg of cloning...

...then be used to infect permissive cells that express ACE2, the cellular receptor for SARS-CoV-2, in a single...

...that was created for systematic screening of extracellular protein interactions to identify novel receptor-ligand...Aggregation Receptor 1 (PEAR1) Ligand. Sun Y et al., Mol Cell Proteomics. 2015 May; 14(5):1265-74. PubMed 25713122...

...Protein Resources: Empty Backbones Biosensors Subcellular localization Optogenetics Background Förster ... Constructs to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses...

...plot of CD3 (T cell marker) versus CD19 (B cell marker) to separate the B cell and T cell into distinct...separate the B cell population into progenitor B cells, immature B cells, plasma cells or others.   ...determining cell health, analyzing cell cycle stages, and detecting specific populations of cells in a complex... the cellular level, it cannot be used for subcellular analysis such as morphology or subcellular localization.... As cells pass through the flow cell the size of the sample stream is reduced forcing the cells to line... photons varies from cell to cell depending on the level of fluorescence. A cell expressing high levels...studies B cells, might then gate the B cell population and create a histogram of CD19-containing cells versus...

...out all CD8+ T cells from a mix of CD8+ T cells, CD4+ T cells, B cells, and dendritic cells. In this case...dumped”. From left to right: B cell, CD4+ T cell, dendritic cell, CD8+ T cell. Created using biorender.com...in flow cytometry are cell proliferation and cell viability. Unlike intracellular labeling, dye staining...be done on live cells. For cellular proliferation, esterified dyes that can enter cells freely are typically...amines on the cell surface, which yields a much weaker signal on live cells compared to dead cells. As a result...comes to labeling cells for flow cytometric analysis, the most common method is a cell surface label, where...through the cell membrane freely, while others require a permeabilized membrane to enter cells. Depending...

...activity, and consists of other cell types, especially immune cells and cells of epithelial origin. Suppressing...turns off T cell function upon interaction with APCs. Furthermore, tumor cells alter their cell-surface proteome...molecules needed for T cell effector function. Cancer cells also produce programmed cell death ligand (PD-L1... activity of CTLs and NK cells. These cells would normally kill cancer cells but are directed not to through...to use whole cell therapeutics consisting of various types of immune cells. Dendritic cells (DCs) are particularly... intravenously. B cells and T cells activated ex vivo are also contestants for cell-based immunotherapy... include: Cell heterogeneity Tumors are composed of a wide variety of cell types. This cell heterogeneity...

...Fluorescent Protein Resources: FRET Biosensors Subcellular localization Michael Davidson Collection Blog...University of Arizona Shaner et al. : Journal of Cell Science, Vol. 120, December 2007, pp. 4247-4260 ...

...Many cell types can be found in the tumor microenvironment (tumor cells, immune cells, blood cells and ...about 53% of my cells are dendritic cells and ~13% are CD8+ T cells. Around 34% of the cells are neither ...neither of the two cell types (likely B cells and others). Almost no cells stain positively for both markers...measure your cells of interest. In many experiments you’ll be working with a heterogeneous cell population...processed piece of tissue, where cells are present that vary in cell type, size, and marker proteins. ...analyzing a homogenous cell population in culture, it is normal to observe diversity in cell size, marker expression...illuminates the passing cell with a laser and detects the light scattered by that cell at a low angle (FSC...

...Pluripotent Stem Cells by Direct Delivery of Reprogramming Proteins." Cell Stem Cell 4, no. 6 (2009): ...Jaenisch. "Induced Pluripotent Stem Cells Meet Genome Editing." Cell Stem Cell 18, no. 5 (2016): 573-86. PubMed... express a cocktail of stem cell reprogramming factors and wait for cells to de-differentiate. However...transgene-free and vector-free cells Genome integration Need to sequence cells to verify excision didn’t ...vector-free cells No genomic integration Can be difficult to fully remove virus from cells Plasmid Non-integrating...transduces a wide range of cell types and replicates in the cytoplasm independent of the cell cycle. A challenge...virus which is removed by culturing cells at 38℃, but screening cells for the presence of transgenes is ...

...performed on whole-cell extracts from the initial stable cell pool (pool) and from individual cell expansions ... in several human cancers) in A549 cells (a human lung cancer cell line), armed only with viruses obtained...need our A549 cells to express Cas9. In these particular experiments, we generated A549 cells stably expressing...52962-LV, Figure 1). Some cells were then used to generate a monoclonal cell lines by limiting dilution...The rest of the cells were frozen down. Figure 1: Cas9 expression in A549 cells transduced with...blotting was performed on whole-cell extracts from stable pools of A549 cells transduced with Cas9 expressing...A549 cells are relatively easy to transduce. I ended up with too many Cas9 integrations per cell, and ...

...levels in your cell lines, you can consider a cell treatment approach. If there is a cell treatment (e....select a candidate cell line (e.g. DepMap) – you’ll need to ensure that the parental cell lines of your chosen...the parental cell line, but not in the iso-genic knockout control. This can involve cell surface labeling...selectivity for cell surface labeling may become less selective if used in intracellular labeling because...because the intracellular compartment may contain cross-reacting antigens not present on the cell surface....looking to label an intracellular target.   Figure 1: HCT 116 WT and SYT1 KO cells were labelled with...) compared to the WT cells (green histogram). The unfilled histograms show cells that have only been stained...

...mycoplasma contamination in your cell culture or you are about to begin a new cell culture project. If mycoplasmas...present in most of the cell culture facilities, tissue culture labs and every cell culturist has to deal...cost-effective (Sung, 2006). The sensor cell-based method uses specialized cells that can detect the presence of... should be made available for cell culturing and maintenance of the cell lines. Providing such an environment...Photo by Kate Harten DeMaio. Cell lines and reagents Examine your cell line stock for mycoplasma contamination...cryopreserved vials of the cell lines even though the viability of cryopreserved cells decreases with every ...cause mycoplasma infection from one cell line to another. Multiple cell lines should not be stored in a single...