...perfect empty backbone, made sure to add the right tags to your gene, and may have even put a fluorescent... your plasmid or carrying out your experiment. History of E. coli strains E. coli are gram-negative, rod... DH5alpha and DH10b (also known as TOP10). The history of B strain is a bit more convoluted due to researchers...researchers sharing and renaming samples throughout history. It was likely isolated in 1918 but was first referred...

...fusing a protein tag to the target and using an epitope-based strategy such as Anti-6xHis. You could also...ICC) when the sample is cultured cells or immunohistochemistry (IHC) with tissues), where an antibody binds....3001161 McLaughlin, K. (2019). Multiplexing Immunohistochemistry. Materials and Methods, 9. https://doi.org...

...Experimentally, antibodies conjugated to fluorescent tags or other markers can be injected into an animal,...antibody and production strategy has advantages and disadvantages compared to each other. Polyclonal antibodies...produced for research purposes, some of which take advantage of natural immune system production and many more...as polyclonal. Polyclonal antibodies have the advantage of more robustly recognizing the target protein...proteins are also plasmid-derived. However, a disadvantage to ScFv is their lower affinity for their target...antigen as canonical antibodies. They also have the advantage of being significantly smaller, meaning they can...proteins you can investigate in each panel. An advantage of flow cytometry is the ability to use multiple...

...PHPeB Clear Filters ID Name Promoter Opsin Variant Tag Activity Serotype PI 20071 pACAGW-ChR2-Venus-AAV ...ChrimsonR (soma-targeted) GCaMP8s Cre dependent 9 Histed 183519 pAAV-CaMKIIa-ChRmine-oScarlet-KV 2.1-WPRE...

... adds a myristoylation sequence and flag-epitope tag to each ORF from the labs of William Hahn and Jean...-A major histocompatibility complex, class I, A FLJ26655, HLAA HLA-B major histocompatibility complex,...major histocompatibility complex, class II, DM beta D6S221E, RING7 HLA-DOA major histocompatibility complex...HLA-DOB major histocompatibility complex, class II, DO beta DOB HLA-DPA1 major histocompatibility complex, ...major histocompatibility complex, class II, DQ alpha 2 HLA-DXA HLA-DQB1 major histocompatibility complex...major histocompatibility complex, class II, DR alpha HLA-DRA1 HLA-DRB1 major histocompatibility complex... HLA-G major histocompatibility complex, class I, G MHC-G HLA-H major histocompatibility complex, class...

.../9R] ZIP3 Mouse Mouse IgG2a 206600 Anti-S-tag [N56/9R] S-tag Bovine Mouse IgG2a 206601 Anti-TrpC4 [N77...206528 Anti-Histone H3 pThr11 [N123/19R] Histone H3 pThr11 Rat Mouse IgG2a 206529 Anti-Histone H3 pThr11...206550 Anti-Synaptotagmin-5 [N274/8R] Synaptotagmin-5 Mouse Mouse IgG2a 206552 Anti-Histone H4-dimethyl-Arg3... Human Mouse IgG2a 114476 Anti-Synaptotagmin-3 [N278/19R] Synaptotagmin-3 Mouse Mouse IgG2a 114477 Anti-TRPML3... Human Mouse IgG2a 114482 Anti-Synaptotagmin-12 [N277/7R] Synaptotagmin-12 Mouse Mouse IgG2a 114483 Anti-GluA1...) Rat Mouse IgG2a 114558 Anti-Synaptotagmin-10 [N269/73R] Synaptotagmin-10 Mouse Mouse IgG2a 114559 Anti-Ataxin...28R] HCN3 Mouse Mouse IgG2a 177500 Anti-6xHis [N144/14R] 6xHis Mouse IgG2a 177501 Anti-TrpV3 [N15/4R] ...

...uracil DNA glycosylase inhibitor and degradation tag on the fusion protein to limit the activity of the...Addgene depositor Luciano Marraffini’s lab took advantage of this lethality to design the first synthetic...the original pCas/pTargetF but has many other advantages including shorter plasmid curing time. pCRISPomyces... al., 2014). These could be genes, specific polymorphisms, including antibiotic resistance genes. This...

...(2017). AAV-mediated Anterograde Transsynaptic Tagging: Mapping Input-Defined Functional Neural Pathways...bodies. Anterograde and retrograde tracing take advantage of existing transport pathways in neurons. Anterograde...by neuroscientists for some time due to their advantageous characteristics for brain research and their...infected cells that also express Cre. Another advantage of AAV vectors is that there are well-established...dye-based tracing while also having the added advantage of lower toxicity, higher specificity and reduced...PubMed Central PMCID: PMC5102064. Roberts, T. F., Hisey, E., Tanaka, M., Kearney, M., Chattree, G., Yang...

...adjusted voltage and compensated your colors. Here are some good practice tips on adjusting voltage: During...changing the voltage for the particular fluorophore prior to recording data. Increasing the voltage will shift...decreasing the voltage will shift it to the left. If your signal is too high, lower the voltage to have your... to see what percentage of total live cells are GFP+. In this plot, with adjusted voltage, cells that ...quantify based on percentage positive or negative in your dot plot, looking at a histogram vs antibody plot...gates, just based on a histogram plot instead of a dot plot. However, the histogram plots look a bit different...that for the flow plots discussed in this post, voltage has been adjusted appropriately and compensation...

...CODA CODA aGTCCGCCTCCACCCGAGGTAGAG zgc:111821 OZ597 and OZ598 CODA CODA tGGCAGCAGCTTTTCGAAGAATAGg zgc:65890...gCGTGACCTCgctgatAAGAAGAGg xylt1 OZ509 and OZ510 OPEN OPEN cACCCTCTACaggacaGTAGCGGGTg zgc:153086 OZ511 and OZ512 OPEN OPEN ...cACCTGCAGCagattGAAGAAGAGg cftr OZ517 and OZ518 OPEN OPEN gTGCAACCGCagctttTAGGACGGAt fox1 (a2bp1l) OZ519 and OZ520 OPEN OPEN...aGACTCCGTCccgtcgtGCTGAGGGCc hnf1a OZ521 and OZ522 OPEN OPEN aACCTTCCCCagagaGAGGTAGTGg VDAC2 OZ523 and OZ524 OPEN OPEN gCTCCCAACCactttGCTGGTGTCa...and OZ548 OPEN OPEN gAGCTCCGTCtcgctgGCGGCCGAAg histamine receptor H3 (hrh3) OZ549 and OZ550 OPEN OPEN gCGCCACGGAcaataaaGAGGACTGCa...tTACTTCTGCaatgatGTGGTGGCTc vipr1 OZ559 and OZ560 OPEN OPEN gGCCATCAGCtcgcattGGAGAAGTAg NRF2b (nfe212b) OZ561 and OZ562 OPEN ...tGTCTCCGGCagaggGAGGAGGAGa esrrga OZ563 and OZ564 CODA CODA tAGCTACCGCTGGCGTCTGAGGAGc HACE1 OZ565 and OZ566 CODA CODA gCTCATCTCCACCGAGCTGGAGAAg...

...apoeb TAL3402 & TAL3403 TGCCAGGCTCGTAGCCTGttccaggctgatgccccTCAGCCCAGATGGGAGGA arap3a TAL3026 & TAL3027...ARSA TAL 3002 & TAL 3003 TCAAGAGGCGGTCTCCCTctcaacgagaccaccaTAGCCGAGGTGCTGAAGA asun TAL3404 & TAL3405 TAACTCAAGCCAACTCCAggaaacatcaggagtttGCAGGAAGATTGAGTTCA...cited2 TAL3228 & TAL3229 TGGTAGACCGCATGATGGcaatgaaccatggtcgTTTCCCAGATGCTGTGAA cited4a TAL3046 & TAL3047...clcn1b TAL3052 & TAL3053 TTTTATTGTGTTACAGACtgcctagcccgtgtccAGAGGTATATTGTGACAA col22A1 TAL3410 & TAL3411...crispld2 TAL3232 & TAL3233 TATCAGGAGGAGCTAGAAcccaacagcactaaacCCGACGCTCCTGTCCGAA cx52.6 TAL3056 & TAL3057...CYP20A1 TAL3414 & TAL3415 TAGACTTTCCTGAAAAGCaaaaatgctagattttGCCATTTTTGCTGTGACA dachc TAL3234 & TAL3235 ...TCCCGTGCAAGGAATGAAtgcaaaaacatttcctTTCCCTTGTTTTGACGGA dcps TAL3416 & TAL3417 TGAACAGTCCACCGATAAcccagtggtagacttcCAGCTCACCCGCATTCTA ddx26b TAL3418 & TAL3419...

...field in which covalent modifications to DNA or histones, the proteins that help package DNA, were associated...modifications, researchers used blunt tools like histone deacetylases, but targeted epigenetic modification...researchers additionally fused a TAL effector to LSD1 histone demethylase to demethylate enhancer regions (Mendenhall...regulate transcription, also includes light-regulated histone methyltransferases and deacetylases. CRISPR and... acetyltransferase increases levels of H3K27ac histone modification at specified loci. Charles Gersbach... Figure 1: dCas9-p300 adds H3K27ac marks to histones in promoter and enhancer regions. These marks ...expression, Fuw-dCas9-Dnmt3a and Fuw-dCas9-Dnmt3a-P2A-tagBFP are available from Rudolf Jaenisch’s lab, with ...

...Coupled Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over...magnetic beads through an assembly of biotinylated SpyTag-SpyCatcher linkers and spacer proteins ending with... single alpha helix) and target capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph...structures (at < 5 Å resolution) of the linker histone H1.8 in Xenopus nucleosomes and in complex with...methods such as chemical crosslinking or tilted-stage imaging. A truncated form of a tardigrade LEA, RvLEAMshort...can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate the endogenous methyltransferase...uses a fusion of the DNMT3L cofactor (D3L) with a histone tail (H3K4me0) to a DNA-binding protein (dCas9 ...

... can be inherited or arise through exposure to mutagens like radiation or viruses (e.g., HPV, the human...TP53 dependent G2 arrest mediator candidate Scotin Shisa family member 5 Sestrins SESN1 SESN2 SESN3 Sestrins...of p300/CBP in p53-dependent signal pathways. Avantaggiati ML, Ogryzko V, Gardner K, Giordano A, Levine...

... genome, it has the potential for insertional mutagensis. The 2nd and 3rd generation lentiviral systems...plasmid 14749 for Thy1.1 selection. Benoist and Mathis 21915 Tet-pLKO-puro 3rd inducible expression of...

...knowledge of protein structure and saturation mutagenesis. mK-GO matures from green to red, and the FT ...absorption spectra). Subach et al’s directed mutagenesis also produced the three separate timers shown....8/4.1 7.1 h (red) pFast-FT-N1 pTRE-Fast-FTpBAD/HisB-Fast-FT Medium-FT 401 (blue), 579 (red) 464 (... 3.9 h (red) pMedium-FT-N1 pTRE-Medium-FT pBAD/HisB-Medium-FT Slow-FT 402 (blue), 583 (red) 465 (...2.6/4.6 28 h (red) pSlow-FT-N1 pTRE-Slow-FT pBAD/HisB-Slow-FT -  Applications of fluorescent protein...

...are sporadic, having no prior family history. A small percentage (5-10%) are familial ALS cases having...

...H2AFX H2AFY H3F3A H3.1 H3.3 HIST1H1E HIST1H2BB HIST2H2BE RBX1 STAG2 Xist Your favorite gene! SWI...context of chromatin?” Though it’s true that adding histones and chromatin architecture to every diagram portraying...transcription of a gene a megabase away, or looking at the histone methylation status at a double-strand break that...2018 Keystone meeting for Chromatin. Note that histone genes frequently have many different variant names...RANbodies) as versatile, small, sensitive immunohistochemical reagents. Yamagata M, Sanes JR Proc Natl ...

... Commons license.  Histograms  For analysis of a single marker, a histogram showing frequency of events...numbers by gated populations; this indicates the percentage of total events contained in the gated population...quadrants to gate and has numbers representing the percentage of cells in each gate. Used under Creative Commons... “true” signal.  Figure 8: A series of histograms from Alyamani et al., 2018, showing number of...

...is caused by a variety of single nucleotide polymorphisms (SNPs) in the TTR gene and is molecularly characterized...targeting the diseased allele specifically. Early-stage trials showed a high success rate in clearing misfolded...above! There are many, many more in the earlier stages of the pipeline, ones that have experienced hiccups...Thus far, several targeted therapies have taken advantage of what our current delivery technologies are ...Plasmids and Resources CRISPR 101 eBook CRISPR History and Development Resources on the Addgene blog...