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We narrowed to 872 results for: cel

Showing: 181 - 200 of 872 results

  1. Antibodies 101: Beyond Surface Labeling

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    Blog Post
    ...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...

  2. Cancer and the Immune System: Deciphering the Relationship

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    ...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...

  3. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...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...

  4. Delivery Methods for Generating iPSCs

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    ...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 ...

  5. Chemogenetics AAV Preps

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    ... chemically induce neuronal activity in specific cell types. See our Chemogenetics Plasmids and Chemogenetics...

  7. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

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    Blog Post
    ...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 ...

  8. Antibody Validation for Flow Cytometry

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    Blog Post
    ...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...

  9. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

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    Blog Post
    ...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...

  10. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...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...experiments including subcellular visualization of 6 different organelles, a cell cycle indicator for mouse...

  11. Plasmids 101: Mammalian Vectors

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    Blog Post
    ...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 ...

  12. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

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    Blog Post
    ...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,...

  13. Antibodies 101: Reading a Flow Plot

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    Blog Post
    ...-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...

  14. Quick Guide to All Things Lentivirus

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    Blog Post
    ...-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...

  15. CRISPR Plasmids - Prime Edit

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    ...and the resulting heteroduplex is resolved by the cell’s mismatch repair (MMR) system. Alternatively, if...

  16. AAVED

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    ...GCaMP6f-P2A-nls-dTomato Detailed protocol: AAV Production in HEK293 Cells Please share materials by uploading to your Dropbox...

  17. Plasmids 101: Protein Expression

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    Blog Post
    ...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 ...

  18. Getting the Most from Your Lentiviral Transduction

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    Blog Post
    ...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...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...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...consumption due to the high cell density of the culture. In addition to benefitting your cells, medium exchanges...

  19. What the HEK?

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    Blog Post
    ...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...

  20. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    Blog Post
    ...producing cells. After FACS, return the cells to doxycycline-free media and expand the cells. Use a subset...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...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...PiggyBac transposase. In rtTA-cell lines, it is better to first engineer the cell line with rtTA, then integrate...own mARGAna cell line. (c) Z-projected phase contrast images of GV expressing MDA-MB-231 cells (top, scalebar...
Showing: 181 - 200 of 872 results