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We narrowed to 819 results for: cell

Showing: 541 - 570 of 819 results

  1. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...immunocytochemistry (ICC) when the sample is cultured cells or immunohistochemistry (IHC) with tissues), where...know if they co-localize at the same place in the cell? Or you know that the presence of several biomarkers...tissue sections, methanol- or paraformaldehyde-fixed cell samples, or other preparations. Sequential IF For...hippocampus, or counting the number of puncta in each cell. When making figures for a paper or presentation...approach for deep phenotyping and spatial analysis of cells in complex tissues. Proceedings of the National ...

  2. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...interest from Addgene, transfected it into your target cells, now what? How can you tell if the protein you are... are so keen to study is expressing in your cells? Immunoblotting or simply the western blot, or western...specific protein within the complex mix derived from cells or tissues. Westerns are useful not only to detect...context (Pillai-Kastoori et al., 2019). Tissues or cell lines other than those used by the vendor for validation...negative controls. Positive controls typically include cell lines or tissues that are known to express the protein...readout for subcellular fractionations, a procedure that isolates proteins from different cellular compartments...protein levels relative to standards, detect the cellular location of proteins, and can be a readout for...

  3. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...desired mutation in mouse ES cells, select for the mutation and inject the ES cells into a mouse embryo to ...Cas-mediated genome engineering." Cell 154 (2013):1370-9. DOI: 10.1016/j.cell.2013.08.022 PubMed: PMID: 23992847...then electroporate it into mouse embryonic stem cells. After selecting for clones with the recombined ... your chimeric mice did not contain edited germ cells, your experiment would be unsuccessful. CRISPR made...

  4. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ...they either trace downstream from the soma (neuron cell body) toward the axon (anterograde tracer) or upstream...particles require receptor-mediated entry into the cell. This means that not all neurons and pathways will...chemogenetic vector directly into the region where the cell bodies of the projection neurons reside.   Impact...

  5. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...co-delivering different AAV particles into the same cells in vivo? A: This will be highly dependent on vector...particularly one that can target the epithelial cells in the blood-brain barrier?   A: For liver, we recommend...worth investigating for both Q6 & Q7.For epithelial cells of the blood brain barrier we would currently recommend...and to all our participants for their time and excellent questions.    Additional Resources on the Addgene...

  6. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ... glutamate release from neurons and other brain cells in vivo. Glutamate plays a variety of roles in synaptic...promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin-1 promoter. Good for neuronal...promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP: Synapsin-1 promoter, FLP-dependent ...sensor to the membrane and displays it on the extracellular side. In addition to the original PDGFR TMD,...

  7. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...electrocompetent cells in which the phage recombination machinery is induced using L-arabinose. These cells can be...Enzymes Required No No Yes Yes Lab Made Competent Cells Required Yes No No No Comparison of REPLACR-mutagenesis...

  8. CRISPR Plasmids - Plants

    Type
    Collection
    ...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 ...Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both...

  9. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ...abundant forms of posttranslational modification in cells and research into its many roles in protein function...interactions can be discovered by fluorescence-activated cell sorting (FACS). The Mode #2 phosphosite library ... production of recombinant phosphoproteins using cell-free protein synthesis . Nature Communications ,...TBK1 activation to promote mitophagy . Molecular Cell , 60 (1), 7–20. https://doi.org/10.1016/j.molcel...

  10. Mammalian RNAi Tools

    Type
    Collection
    ... in therapeutic settings. As a tool in mammalian cell systems, silencing is achieved through the delivery...). Origins and mechanisms of miRNAs and siRNAs . Cell, 136 (4), 642–655. PubMed (Link opens in a new window...

  11. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...high-throughput screening techniques such as ELISA and cell surface display are common. ELISA enables quantitative...for comparing relative affinities across mutants. Cell surface display systems, such as bacterial, yeast...presentation of your mutant libraries on the surface of cells. This enables direct screening via flow cytometry...provide recombinant antibodies that target families of cell surface and secreted proteins (glypicans, integrins...therapies and improve their delivery methods to various cells. The wealth of smaller engineered antibody fragments...

  12. Tips for Getting a Faculty Position

    Type
    Blog Post
    ...patterning of cells and tissues is fundamental to development. I'm interested in how cells find their way...competitive CV You don't necessarily need a Nature/Cell/Science paper. You do need multiple first/corresponding... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

  15. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...Mutations. Berger AH, Brooks AN, Wu X, et al. Cancer Cell, 2016. PubMed PMID: 27478040. The lung cancer mutant... profiling of human gene and allele function via Cell Painting. Rohban MH, Elife, 2017. PubMed PMID: 28315521... of MAPK1/ERK2 Missense Mutants. Brenan L et al. Cell Reports, 2016. PubMed PMID: 27760319. PubMed Central...poorly classified function and a well established cellular signaling gene, thus providing a starting point...reference cDNA clones mapping to well-annotated cellular pathways). It wasn’t clear whether gene-agnostic...sequence-confirmed series of clones corresponding to known cellular signaling pathways as part of our Genetic Pathway...functional assays for these mutations and map other cellular functions and phenotypes. Since time is of the...

  16. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...toolkit in human cells. Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK. Mol Cell. 2014 May 22;54(...tRNAs are among the most abundant molecules in a cell, this system is clearly also robust enough to ensure...advantageous to instead use a preexisting, conserved cellular machinery to process gRNAs. Hijacking tRNA processing...

  17. How Exercise Purges Stress and Keeps Depression at Bay

    Type
    Blog Post
    ... how it works. The findings in a recent issue of Cell demonstrate how PGC-1a1, a transcriptional coactivator... in a commentary about the new work published in Cell Metabolism. But, they note, there is plenty more...JR, Erhardt S, Lindskog M, Ruas JL. doi: 10.1016/j.cell.2014.07.051. PMID: 25259918 Muscle over mind. Moon...

  18. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ... For most kits, the steps for lysing cells and removing cellular debris look very similar. However, keep...relative term — even armed with the highest-competency cells you will still struggle to get transformants if ...quantity) – Depending on the experiment, mammalian cell transfections can require quite large amounts of...your bacteria to release the plasmid, separate the cell debris from the plasmid, and then clean-up the plasmid...Kits can be optimized based on the plasmid size, cell model, or overall throughput needed. Large constructs...your plasmid; separate plasmid DNA from unwanted cellular “debris”; and then clean-up the plasmid DNA (Figure...

  19. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...with heterotrimeric G proteins on the intracellular side of the cell membrane. Heterotrimeric G proteins...Peroxidase-Catalyzed Proximity Labeling. Cell 169:338-349.e11 . https://doi.org/10.1016/j.cell.2017.03.028 Roth BL (2016...small molecules to elicit a response in a certain cell type. Chemogenetics is similar to optogenetics but...induce or inhibit intracellular signaling cascades as a response to the extracellular stimuli. Ligand dissociation...to the seven alpha helices that transverse the cellular membrane. The receptor recognizes the appropriate...signaling is initiated when a ligand binds to the extracellular surface of the GPCR. This results in a conformational...

  20. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Additional resources... also detect low-frequency cancer mutations from cell-free DNA fragments as well as health-related single...similar types of HPV, HPV16 and HPV18, from human cells, at atomolar levels, within one hour. DETECTR thus...of Diverse Class 2 CRISPR-Cas Systems. Molecular Cell 60:385–397 . https://doi.org/10.1016/j.molcel.2015.10.008...

  21. What's New in CRISPR - December 2019

    Type
    Blog Post
    ...mixture of two base editor enzymes in mammalian cells. Read the paper in bioRxiv Find Target-ACE   CRISPR...bionanoparticle Transient expression of CRISPR/Cas9 in cells has advantages like reduced off-target activity ...

  22. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

    Type
    Blog Post
    ... brain regions. You can partially control which cells are successfully infected by an AAV by altering ...control where your gene of interest is expressed.  A cell-type specific Cre can provide you with that control...

  23. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    ...multiple plasmids in one cell, it might be that you can't stably maintain them in the cell. This blog post features...

  24. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    ...pathways that are frequently implicated in cancer cell proliferation, survival, differentiation, and apoptosis...pathway. “The idea is that if you introduce them into cells one at a time, you could identify those that confer...

  25. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... GFP, enabling you to select cells with high GFP expression. These cells have the highest levels of Cas9... on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the desired...gRNA, which causes hypopigmentation, thus marking cells that have undergone genomic modification. In this..., et al. “Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system.” Scientific...

  26. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...Drosophilia Plasmid pRDB_051 (for stable fly cell lines; attL and attR sequences for genome integration...Drosophilia Plasmid pRDB_052 (for stable fly cell lines; Hygromycin resistance gene; for genome integration...rapid assessment of emerging CRISPR technologies. Cell Genomics, 4(3), 100519. https://doi.org/10.1016/...

  27. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...lack uracil, but contain methionine. (This allows cells carrying nonfunctional as well as functional MET... is available at our Investigations in Molecular Cell Biology Website. Many thanks to our guest blogger...right) has taught courses in biochemistry, molecular cell biology and genetics at Boston College. She is particularly...

  28. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...currently doing this using two approaches: single-cell RNA-sequencing to identify neuronal class and deep...WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans embryo when all essential...Wang et al., 2018).” With the split system, only cells that have both halves of cGAL can drive expression...

  29. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ... monitoring sensitized emission in living plant cells." (2013). PubMed PMID: 24194740. PubMed Central ...-molecule FRET for imaging biomolecules in live cells (Sustarsic and Kapanidis 2015), which may lead to...understanding biomolecular structure and dynamics in live cells." Current opinion in structural biology 34 (2015...fluorescent proteins available. Fortunately, several excellent reviews have collated data from various papers... alters the FRET efficiency. Detecting certain cellular changes with an intermolecular probe is often ...Miyawaki et al., 1997), was designed to measure intracellular calcium and published in 1997. Since that time...of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments can probe...

  30. Editor's Choice, August 2016

    Type
    Blog Post
    ...or G->A transitions in a target sequence in live cells. This technique uses catalytically inactive CRISPR...and other means of generating mutations in live cells are all discussed further in this informative post...Clearly we have all of our writers to thank for the excellent content they’ve churned out in the last month....
Showing: 541 - 570 of 819 results