We narrowed to 1,027 results for: ELL

... the plasmids of interest (for example, fewer satellite colonies will grow). However, it is also more ...

...homology. While this approach doesn't work in other cell types, it can be a major time- and cost-saver for...

...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...compatible with 96-well plates as collection vessels and can sort single cells into each well allowing users...

...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...

...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...Reprogram Peripheral Blood Cells Check Out Our Interview with Stem Cell Researcher Darrell Kotton Four Factors... 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...

...label an intracellular target.   Figure 1: HCT 116 WT and SYT1 KO cells were labelled with a green...levels in your cell lines, you can consider a cell treatment approach. If there is a cell treatment (e....antibody shown to perform well in one set of conditions may not perform as well in other conditions. For...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....

...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 ...

...experiments including subcellular visualization of 6 different organelles, a cell cycle indicator for mouse...recorded in the cell's DNA under the right conditions, and could initiate specified cellular processes or...Understanding intracellular pH regulation is important as pH regulation has roles in many cellular functions...signaling pathways to the cell cycle. Good tools for measuring intracellular pH are required in order ...useful tools for light-mediated subcellular localization in mammalian cell culture and reversible control...in live cells is that one can now follow movement of two or more targeted loci during the cell cycle or...mammalian cells, yeast, and Drosophila. Synchronization of secretory protein traffic in mammalian cells The...

...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...

...by source cell based on single-cell RNA-seq data. In addition to unique proteins on each cell type, the...regions, cell types, and tissues. Find cell-surface HRP plasmids here! Wu, L., et al. (2025) The cell-surface...this post: Cell-surface protein labeling tools for neurons and astrocytes Nerve-to-cancer cell mitochondrial...AAV-CRISPRa-engineered fat cells fight cancer Frugal CRISPR kit for classrooms Cell-surface proteomics for...for neurons and astrocytes By Mike Lacy Cell-cell interactions underlie many kinds of tissue function,...mapped their cell-surface proteomes (Wu et al., 2025). The team expressed extracellularly-anchored horseradish...the cell-surface shared proteome of astrocytes and neurons (CS SPAN) featured extracellular matrix proteins...

...-B+ cells, and A+B+ cells, but no cells that are A+B-.      Figure 3: It's often easier to identify...population of B cells and dendritic cells (DCs) in the lower middle right. DCs and B cells are HLA-DR+ and...single cells in a population. It can be used to measure protein expression, identify rare cells, or even...looking for cells in a sample that are negative for marker A and positive for marker B, or A-B+ cells. If you...the cell populations as a guide  (Figure 3.) You can see that in our dot plot, we have A-B- cells, A-B...the first step is to separate cells from debris and roughly group the cells by size. This is done using ... and FSC are measured to differentiate cells from debris. Cells are gated, and in the gated population...

...plasmids for RapID EXCELL: Proximity labeling for the detection of cell-cell interactions EXCELL is an enzyme-mediated...contexts.  Proximity labelling relies on the labelling of neighboring proteins within a cell with biotin. Biotin...enzyme-mediated proximity cell labeling strategy to detect cell-cell interactions in vivo (Ge et al., 2019...et al. "Enzyme-Mediated Intercellular Proximity Labeling for Detecting Cell–Cell Interactions." Journal...avidin, proximity labelling can capture weak/transient protein interactions as well as be used for both...a bait protein fused to BirA* in cells and then incubate those cells with exogenous biotin. After a few...BioID and APEX have been developed as well as new proximity labelling techniques. These newer tools are also...

...-dividing cells. The ability to infect non-dividing cells is not restricted to in vitro cell culture as...target cells. If you are transducing non-adherent cells you might need to centrifuge your cells with the...favourite gene into your favourite cell line or into primary cells, this blog article will give you some...infect both dividing and non-dividing cells, including stem cells. These properties make lentiviral vectors...immortalized cells lines but reach their limits when it comes to transferring DNA into primary cells or non-...non-dividing cells. To target these un-transfectable cells, labs use viral vectors and particularly retroviral...will first need to transfect producing cells, usually 293T cells, with your transfer plasmid and your packaging...

...produced using complex mixtures derived from cells or using live cells. There are several types of expression...consists of transfecting cells with your DNA template of choice and allowing these cells to transcribe, translate...demanding cell culture conditions compared to other systems. Insect expression systems Insect cells can also...coli or yeast cells. The only problem with baculovirus systems is that the infected host cell is eventually...eventually lysed. Cell lysis halts protein production, but there are non-lytic insect cell expression systems...Sf21, Hi-5 cells)  that allow for continuous expression of genes integrated into the insect cell genome. ...that is then transformed into a bacterial cell. Transformed cells propagate, are induced to produce your ...

... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ...plasmid to generate a population of mutant cells. The mutant cells were treated with WNV at a dose and duration...regulate the ability of WNV to induce cell death in infected cells (SEL1L, UBE2J1, EMC3, EMC2, DERL2, UBE2G2...screens in human cells. General information regarding how to use a CRISPR pooled library as well as criteria...small GTPases involved in cell signaling pathways that control many cellular processes such as differentiation...community as well. The pooled library, designed to be used with cultured Drosophila cells, targets 13,...and analysing autophagy of cytosol and organelles in yeast cells. Named after the brightly-coloured Australian...

...transfected cells may be considered an entirely new cell line from that of the original parental cells. Positive...cultured mammalian cells. Of course, these mammalian vectors must be compatible with the cell type they are...bacterial cells); however, the techniques and reagents vary. Plasmid transfection into mammalian cells is fairly...expression levels in the cells. Ideally, the chosen reporter is unique to the cell, is expressed from the...components expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr...mammalian cells To achieve stable transfection, there should be a selective pressure to force cells to incorporate...(basically growing cells in various concentrations of the selection reagent). Cells should die within ...

...and spread of cancer cells. Besides these stromal cell modifications, stromal cells are also constantly...how cancer cells influence microenvironments and vice versa, let’s take a look at cancer cell metabolism...inflammation, a noteworthy cell type in the tumor microenvironment is the immune cells. The contradiction is...injecting breast cancer cells into mice, exosomes expressed specific integrins (cell-adhesion proteins) targeting... be transferred to other cells, inducing epigenetic changes in targeted cells. In this case, exosomes ...Friendly” microenvironment cells Not all is bad news – we also have “friendly” cells in microenvironments! ...Take myoepithelial cells for instance. They retard tumor formation by inhibiting cell growth, invasion,...

...on or in these cells: antibodies can bind to proteins on the cell surface, or the cell membrane can be...same cell simultaneously as well as compare between samples. Learn more about flow cytometry. Cell and ...antibodies are produced by B cells. They bind to proteins on the surface of extracellular pathogens such as parasites...fusing B cells that have been extracted from animals injected with protein antigen to myeloma cells. The ... measurement of protein expression in cell lysates, whole cells, or tissue samples. Many experiments that...shows that a similar concentration of cell lysate was added to each well. Image from Sun et al., 2010. ...heterogeneous mixture - for example, cell lysate or media from cultured cells. ELISA can be used to quantitatively...

...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...therefore titers obtained in one cell type may not translate well to other cell types. To overcome this issue...Love your cells and they'll love you back A successful lentiviral infection begins with your cells, both ...both your packaging cells and your target cells. Poor tissue culture practices can lead to low titers and... promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent...reaching 108 cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients... your producer cells with fresh medium. Daily medium exchanges help alleviate cellular stresses associated...

...producing cells. After FACS, return the cells to doxycycline-free media and expand the cells. Use a subset...CMV-gvpNJKFGWV plasmid into 70%-confluent well of HEK cells in a 6-well dish and change the media every day ...white inclusions in cells with GVs via phase contrast microscopy, GVs visible in cells via TEM, and purified... heterogeneity required single cell screening to isolate a clonal cell line capable of producing ultrasound...able to express Anabaena GVs in mammalian cells without the shell stiffening protein GvpC, which allowed ...expression homogeneity, as the cells could be batch-sorted to isolate a polyclonal cell line with strong gas vesicle...transient transfection in rtTA+ cell lines (such as HEK293-TETOn cells), or they can also be integrated...

... your cell type of interest, while miRNA target sites can be used to prevent expression in cell types ... promoter/enhancer strengths in the cell type of interest as well as the type of transgene delivered: ...endothelial cells in rodent CNS AAV-PHP.V1 shows higher transduction in CNS endothelial cells (60%) and ...endothelial cells in the retina and spinal cord (Krolak et al., 2022). Systemic capsids for broad cell type ...delivery across species. Cell, 184(19), 4919-4938.e22. https://doi.org/10.1016/j.cell.2021.08.028 Vormstein-Schneider...system (PNS). (Capsids, if you recall, are the outer shell of an AAV vector, which determine the vector’s tropism... discuss how systemic capsids are engineered, as well as highlight some considerations for their use. ...

... cloned into E. coli cells, isolated colonies will be lifted onto nitrocellulose membranes and exposed... proteins directly onto the nitrocellulose for subsequent probing for cell-binding activity.  First, cut...with live host cells Now you can proceed with incubating this membrane with your cells of interest and...to bind their target cells, pulling them out of solution. Wash your membranes well with PBST to remove ...remove unbound cells. To detect bound cells, simply lay each membrane on an agar plate selective for your target...appropriate cell growth conditions. Note: if using fast-growing bacteria like E. coli or Salmonella, or any...any cells that bind strongly to nitrocellulose, over-incubation of plates can lead to high background that...

...embryonic kidney (HEK) cells originated from? Or why they are such a commonly used cell culture model? In ...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first ...inconsistent with kidney cells (Shaw et al.). Since then, transcriptome studies of the cell line have determined...for?   As one of the older cell lines in the research community, many cellular research tools have been ...any unwanted material (HEK cells, etc.) and are ready to use on another cell or organism. This pipeline...where HEK cells may be particularly useful due to their intrinsic properties. HEK-293 cells are tumorigenic...researchers using the cell line. What the HEK is wrong with it?   All cultured human cell lines are susceptible...

... subset of T-cells, but it is unclear if enough T-cells could be reached to ablate a well-established ...infected cells, the other using wild-type Cas9 to remove the HIV-1 genome from infected cells. Read on...1 reservoirs HIV-1 infects cells in the immune system, notably CD4+ T-cells, and eventually leads to acquired...promoter-luciferase constructs. In multiple HIV-1 latent T cell lines, as well as a latent microglial line, treatment with...percentage of cells with activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that... genome from every infected cell and 2. prevent reinfection of those cells. Kaminski et al. designed gRNAs...Kaminski et al. observed no negative effects on cell viability, cell cycle progression, or apoptosis with Cas9...

...target cells; you also have to consider how to keep it there. In fast-growing populations of cells, the ...into the host cell’s genome, so no special sequence is needed to retain the vector as cells divide. In transient...which helps increase cellular uptake, but this approach requires compatible cell lines that express viral...Alternatively, some slow-growing cells, particularly mammalian cells like neurons, may not replicate during...transfect the vector a viral ori in a compatible cell line cells that replicate slower than the duration of ...is delivered to your target cells, you still need to select for the cells that contain it. Antibiotic ...penicillin and ampicillin inhibit cell wall synthesis. Eukaryotes, with no cell walls, can grow alongside these...

...human embryonic stem cells. Transduced stem cells were differentiated and single cell RNA-seq was performed...transduced human embryonic stem cells with the viral TF library, with each cell receiving only 1 TF expression...construct. Cells were cultured following a differentiation protocol before performing single-cell RNA sequencing... 206 cells per TF ORF. From this large sequencing endeavor, a map of possible TF-induced cellular states...differentiation.   Fig. 2: Single cell RNA-seq clustering of cells transduced with the TF pooled library... clusters of individual cells along with color coded nominated reference cell types.  (Adapted from Joung...delivered to each cell, future applications could deliver multiple TFs to each cell, allowing for combinatorial...

...determine cell cycle length by examining the ratio of blue to red cells. Cells undergoing rapid cell cycles...engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular protein. They used...’s difficult to observe the cell cycle in vivo, particularly if the cells are migratory or in deep tissues... BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations of IL-3. Cells grown...proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in media without IL-3 show...H2B-FT consistently predicted the cell cycle speed across these different cell types. In one example, H2B-FT...expressed in murine B cells (BaF3), which depend on IL-3 to proliferate. The cells cultured with a low ...

... target to specific subcellular regions, laser ablation can damage other cellular structures, and genetic...demonstrated its use to probe cell mechanics, vesicle transport, and cell division, but given the long ...GTPase, leading to behaviors like lamellipodia or filopodia formation, cell contraction, or other morphological... expressed in a monolayer of endothelial cells, showing cell protrusions and overlaps (arrows) after blue...2016). Rho GTPase signalling in cell migration. Current Opinion in Cell Biology, 36, 103–112. https://doi.org...Optogenetics gives you the power to control cells or organisms with the flip of a switch. You may be ...optogenetic tools to control the cytoskeleton and cell membrane dynamics. First, a quick refresher.  The...