We narrowed to 668 results for: cell

...transformation efficiency of the strain: when asking the cell to perform multiple recombination events, all linear...linear fragments need to get into the same cell. For simple cloning requiring just one or two recombination...recombination events, homemade competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications...modifications will require ultracompetent (109 CFU/μg) cells. Primer design Using specific primer design (Figure...

...using an electric charge to help the DNA enter the cells. Some samples require a reduced antibiotic concentration...concentration or need to be transformed into different cells than were originally requested. Most of our samples... Start by scanning the sample barcodes into the cells in the same order that they exist within the rack...storage process - from deposited DNA to transformed cells and finally to bacterial glycerol stocks that are...

...not generally in a cell’s best interest to overexpress a protein. Energy and cellular resources are being...being spent to make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes...proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical and...same manner as it would have in the context of a cell. FLAG tags can be cleaved by enterokinase, which...

...selection markers (for use in other cell types, e.g. eukaryotic cells). pBACKBONE-XXXXX Tip: For a catalog...

...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...

...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 ...

...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...

...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...

...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...

... 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,...

...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...

...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...

...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...

...plasmids is associated with the "Barcoding cells using cell-surface programmable DNA-binding domains" ...

...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...

...bacteria, so it can be transformed as-is into cells. The cells then get to work fixing up the “damage” and...nicked plasmid together long enough to get it into cells and let them do the repair work. The key is T4 DNA...

...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...

... 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...

... 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...

...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...