...transcription factors such as PAX7, PAX6, AP-2 alpha and ISLET-1. In addition, we distribute monoclonal antibodies...Monoclonal Antibody Research Institute dedicated to: 1) developing new ways of generating antibodies, for...proteins of interest for labeling, quantification, purification, chromatin immunoprecipitation and more. The...

... We now have another model of Cas9 – a flexible alpha-carbon backbone model made of nylon – that more ... – a mechanical device that could be used to bend 1/8th inch steel wire into a backbone structure of a...BioMolecular Modeling (CBM) in 1999 with the expressed purpose of creating physical models of proteins and other...

...10.1038/s41551-024-01227-1. doi:  https://doi.org/10.1038/s41551-024-01227-1   CHARM: A compact epigenetic...with a GFP nanobody (Figure 1). This enrichment streamlines the purification and minimizes sample loss ...peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm single alpha helix) and target capture module (SpyTag-GFP nanobody...highly-visible beads on the cryo-EM grid.     Figure 1: MagIC-cryo-EM for sample enrichment and structure...single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking...protocols typically require high concentration and purity of the target molecule, which can be problematic...

...successfully generated several functional heterotrimeric G-alpha subunit fusions that do not tolerate N- and C-terminal... remains fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained in...follow him on twitter: @joachimgoedhart.   References 1. Zacharias, David A., et al. "Partitioning of lipid-modified...vitro by measuring the fluorescence intensity of purified protein at different acid concentrations. Overviews...

...Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR for targeted...well as pLV-dCas9-p300-P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds H3K27ac marks...lentiviral transduction. Lysine-specific Demethylase 1 (LSD1) Tatjana Sauka-Spengler's lab has deposited ...CRISPR, and Addgene has a number of tools for this purpose. Epigenetics began as a correlative field in which...mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced cells..., Korać P, Julg B, Klasić M, Zoldoš V (2016) Repurposing the CRISPR-Cas9 system for targeted DNA methylation...

...Thorel, F., et al., Conversion of adult pancreatic alpha-cells to beta-cells after extreme beta-cell loss...type of choice in vitro is known as reprogramming [1]. The process can be divided into two stages: Dedifferentiation...//www.linkedin.com/in/kmukherjeephd/.  References 1. Hochedlinger, K. and R. Jaenisch, Nuclear reprogramming...from human peripheral blood. Cell Stem Cell, 2010. 7(1): p. 15-9. PubMed PMID: 20621044. PubMed Central PMCID...induced pluripotent stem cells. Hepatology, 2010. 51(1): p. 297-305. PubMed PMID: 19998274. PubMed Central... pluripotent stem cells. Nat Biotechnol, 2014. 32(1): p. 84-91. PubMed PMID: 24291815. PubMed Central ...keratinocyte lineage. Methods Mol Biol, 2014. 1195: p. 1-12. PubMed PMID: 24510784. PubMed Central PMCID: PMC4096605...

...been used for a variety of experimental purposes including (1) single or multiple, constitutive or inducible...Rtn4a-GFP (tubular ER) Microtubules mCherry-alpha-tubulin Early Endosome Vacuolar Compartment...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...

...disadvantages: Table 1 (Joglekar et al., 2017)  Table 1 (Gutierrez-Guerrero et al., 2020) Table 1 (Cronin et al...polyA tail.   Figure 1: Lentiviral production uses three plasmids: (1) The transfer plasmid, (2)...that can be used for specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al, 2020...in cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars were less stable than viruses...express these sugars. (Takeuchi et al., 1997). The alphaGal sugars end up in the envelope and are targets ...targets for complement-based killing by anti-alphaGal antibodies. The VSV-G envelope protein is inactivated ..., and spumaviruses to human serum by galactosyl(alpha1-3)galactosylation. Journal of virology 71:6174–...

...Table 1 below outlines a few of the more common genetic changes found in E. coli strains. Table 1: Common...amber (UAG) stop codon by tyrosine insertion λ-thi-1 or thi1 Mutation in thiamine metabolism Requires exogenous... all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain Natural...plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF...) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109   General cloning and...strain for cloning repetitive DNA. endA1 glnV44 thi-1 relA1 gyrA96 recA1 mcrB+ Δ(lac-proAB) e14- [F' traD36...of an E. coli K-12 strain. F- λ- ilvG- rfb-50 rph-1 NEB Stable   For cloning into and storage of lentiviral...

...-fDIO mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg...pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 N protein  pLVX-EF1alpha-SARS-CoV...2xStrep-IRES-Puro: Expression of the SARS-CoV-2 M protein pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro: Expression...the E protein (pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) and NSP2 (pLVX-EF1alpha-SARS-CoV-2-nsp2-2xStrep-IRES-Puro...2xStrep-IRES-Puro) are also in the works in and should be come available in the next few weeks. CRISPRi...

...you can transform 1-2μl of your ligation reaction into competent cells such as DH5alpha or TOP10. If using...well, so let’s go over the details step-by-step. 1. Digestion Set up restriction digests for your insert...recipient plasmid : insert ratio” of approximately 1:3. Since the number of base pairs for each varies,...at minimum, two transformations after a ligation: 1. Control Transformation containing the ligation mixture...result in few colonies on both plates (unsuccessful 1), in a vector alone plate with many more colonies ...them. An easy way to do this is gel purification. In gel purification, you use a voltage difference across...the eyes and skin. When running a gel for purification purposes it is important to have nice crisp bands...

...into account, the digest pattern depicted in Lane 1 would be expected for a XbaI/NotI digest. The program... these enzymes naturally occur in bacteria for purposes other than cloning or confirming plasmids. It ...cleave DNA. Common lab E. coli K12 strains such as DH5alpha contain 3 methylases that recognize and methylate...will cut? Whether performing a digest for cloning purposes or for diagnostics, we suggest double checking...–/dcm– strain of E. coli such as JM110 and re-purifying it. These specialized E. coli strains have been...

...faster. Years of science, done in weeks Figure 1: Sending out 80 FedEx envelopes in a single day --... 2xStrep-tag for easy detection and affinity purification. All but one are in a pLVX vector background... if you need to. Additionally, they have an IRES-Puro marker to facilitate stable cell clone selection...selection. Expression is driven by a fairly strong EF1alpha promoter. We have verified expression of correctly-sized...

...instead conveniently provide them on your smart phone. 1. Lab Archives by LabArchives The single, most important... life is the lab notebook. Lab Archives is a multipurpose application that allows you to share your data...restriction site to a primer, you only need to skim the alphabet on the side of the screen to find the sequence...

...levels of DnaA to get the job done.   Figure 1: DNA melting by DnaA and IHF is the first step of ...plasmids may result in low yields during DNA purification. On the other hand, they also impose a lower... Mizobuchi, K. (1998). Copy number control of IncIalpha plasmid ColIb-P9 by competition between pseudoknot...