We narrowed to 883 results for: prot

...antibody. Is this protein or protein region highly conserved across species? If the proteins are highly homologous...work very well for applications that denature a protein to its primary (linear) structure using heat or... an antibody in an assay for a native (folded) protein, such as immunoprecipitation, the epitope may not... trying to visualize the precise location of a protein in a tissue section then a directly-conjugated ...conjugates complex. If you are trying to visualize a protein in low abundance on a Western blot, a secondary...antibody that has been validated in the species your protein of interest comes from. If one is not available...different type of antibody and therefore each target protein. This can be accomplished by using a panel of primary...

...Photoactivatable Fluorescent Proteins Control Protein Expression with Cre-lox Control Protein Activity with GFP ... other kinases and proteins. Zhou et al suggest that more photoswitchable proteins could potentially be...transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to the next members of... by inserting pdDronpa domains into loops of a protein of interest. If you plan to try this, it’s important...important to check that the function of the protein and pdDronpa are preserved. References 1. Zhou, Xin...reporter plasmids Find photoactivatable fluorescent proteins in empty backbones in the Michael Davidson Collection...Photoswitchable Fluorescent Proteins ...

...chemically competent cells (protocol) Preparation of electrocompetent cells (protocol) At Addgene, we use...using the protocols provided above, then you can find the subsequent transformation protocols below: Untergasser...chemically competent cells (protocol)  Transformation of electrocompetent cells (protocol)  Cells prepared ...streamlining the transformation step. Read on to find protocols and tips that you can use in your own lab. How...competent cells will work just fine.  A number of protocols for making competent cells are available online...can be transformed without heat shock. Detailed protocols are available via Zymo Research. In brief, we ... us less than 5 cents per transformation. This protocol has worked well for us for multiple E. coli strains...

...caused by Arg-mutant proteins, but there is currently no evidence of Arg-mutant protein rescue in living ...Arg-mutant proteins in living baker’s yeast cells, including Arg-mutant yeast homologs of human proteins that... his lab’s research is the mechanisms by which proteins assemble into functional multi-subunit macromolecular...rare diseases. Spontaneous mutations that change protein sequence affect arginine (Arg) amino acids more...function of even a small fraction of Arg-mutant proteins could improve health outcomes for millions of ...large number of other disease-linked Arg-mutant proteins both in yeast and human cells in order to determine...rescue of a variety of disease-linked Arg-mutant proteins in the lab, clinicians might consider prescribing...

...types of proteins: fiber, penton and hexon proteins. The viral genome DNA is further protected by histone-like...histone-like proteins forming the nucleocapsid. B) Three generations of adenoviral vectors: the first generation...helper adenovirus is required to provide the viral proteins in trans. The Gutless AdV have generated high ... systems are still being developed. A detailed protocol for production of Gutless AdV can be found in ...by looking at the ration of DNA (260nm) versus protein (280nm) absorbance. The ratio for the purified ... Enjoy! References 1. Luo, Jinyong, et al. "A protocol for rapid generation of recombinant adenoviruses...adenoviruses using the AdEasy system." Nature protocols 2.5 (2007): 1236. PubMed PMID: 17546019. 2. He TC. et ...

... process. When using zinc finger proteins or TALE proteins, proteins had to be re-engineered for each ...VP64 fusion protein engineered with aptamers that bind to MS2 proteins. These MS2 proteins then recruit...simplicity of gene activation: rather than requiring protein engineering for each loci, CRISPR/Cas systems only...activation CRISPR activation uses dCas9, a CRISPR protein variant lacking its endonuclease ability, to bind...transcriptional activators: some methods rely on fusion proteins while others re-engineer components of Cas systems...CRISPR activators is that it requires a fusion protein, rather than relying on a two-component system ...Gilbert LA, Qi LS, Weissman JS, Vale RD (2014) A Protein-Tagging System for Signal Amplification in Gene...

...It was a bit intimidating, but I had my lab’s protocols and some great co-workers to help me out. Even...writing AAV production tricks into the margins of my protocol. While these tips weren’t critical to the experiment...replication uses the host’s polymerase, but requires Rep proteins to process a double-stranded intermediate into...single-stranded genome that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper plasmid which ...HEK293 cells. Check out the Addgene AAV Production protocol for more details. Pro tips I’m not going to lie...Stable. NEB Stable competent cells lack RecA, a protein that aids in the homologous recombination of duplicated...enough AAV. Compared to the Addgene AAV Production protocol, I take a lazier approach for harvesting my AAV...

...yellow fluorescent protein mGold2 By Mike Lacy Get your sunglasses — these proteins are bright! Most current...post: Bright and photostable yellow fluorescent protein mGold2 Viral delivery of TnpB for plant genome ... current yellow fluorescent proteins tend to photobleach rapidly, which hinders their use for long-term...recently developed a pair of new yellow fluorescent proteins: mGold2s and mGold2t (Lee & Lai, et al., 2025)...or mCitrine, or if existing yellow fluorescent proteins just haven’t met your needs, consider trying mGold2s...2025). Bright and photostable yellow fluorescent proteins for extended imaging. Nature Communications, 16...production processes. Check out our step-by-step protocols for AAV production, purification, and titration...

...recapitulate the results of Gao et al 2016 in a letter to Protein & Cell. Update (August 3rd, 2017) THE ORIGINAL ...activity of the enzyme. In vivo, only nascent NgAgo protein (probably while it is being synthesized) can form...section asked basic questions about the NgAgo protocols used by respondents (transfection method, amount... to wait until someone else optimizes an NgAgo protocol for their favorite model system. The vast majority...trickiness of NgAgo is the inability of mature NgAgo protein to load an ssDNA guide. It is possible that, during...translation, only a small fraction of nascent NgAgo protein molecules manages to form a functional complex ...system. I encourage you to share your results and protocols with the research community if you've had success...

...Green Fluorescent Protein Resources on Addgene.org Browse our Fluorescent Protein Collection Find Plasmid...sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging technologies...technologies, the combination of multiple fluorescent protein sequences to develop diverse fluorescent reporters... reporters can be used to detect structure and protein localization, as well as cellular processes like...modules used in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals... created, each containing a unique fluorescent protein with minimal spectral overlap between them (Table...In each construct, 3 copies of the fluorescent protein were fused in-frame to a tethering partner, which...

...neuroscience-related proteins, but we recently added several broadly useful antibodies for detecting protein tags.  ... saturating mutagenesis on 21 positions in the protein sequence of the predecessor GEVIs (called ASAP ...Kalium channelrhodopsins (KCRs), from a fungus-like protist Hyphochytrium catenoides (Govorunova et al. 2022...  And when you think of protein tags, what are the first tags that come to mind? I’m willing to guess ...EQKLISEEDL, that can be incorporated into your protein of interest and recognized by reliable antibodies...derived from the C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first...Myc tag has become one of the most commonly used protein tags - a quick search for ‘Myc tag’ in the Addgene...

...paper or protocol. If you have questions while reading these materials (maybe a step in the protocol you don...ask for a copy of the protocol if you prefer to take notes on a printed protocol, or maybe you would like...resources to help you get started!) Are there protocols you can familiarize yourself with? If your bench...bench mentor hasn’t sent you papers or protocols yet, don’t be scared to ask for some! You can email or...or “It would be great if you could send me the protocol we’ll be using next week so I can familiarize ...bare minimum should be taking notes on general protocol steps and other tips and tricks your mentor shares...next few days or weeks), look over your notes and protocol(s). Is there enough information there for you ...

...such as Rep or other initiator proteins, they do not require host proteins for replication and can maintain...as recognition sites for replication initiation proteins. These iterons vary in their sequence and number...oris interact with various RNA polymerases and proteins from bacterial hosts. In a plasmid with multiple... (ori1, ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various...has its own replication initiation factors and proteins independent of the host. Host range depends on...can replicate in different bacteria and express proteins within different host organisms. pBBR1-based plasmids...Pasteurella multocida, Porphyromonas gingivalis, Proteus mirabilis, Providencia spp., Pseudomonas spp., ...

...fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level....system to express one of the three fluorescent protein (XFP) genes in a transgene. When multiple copies...2018) (Figure 1). Once introduced, fluorescent proteins were best imaged after tissue clearing with SeeDB2.... For more details on this procedure, find the protocol here.   Tips and troubleshooting your Tetbow...expression levels. Clearing large tissues - Fluorescent proteins are very stable in SeeDB2. However, SeeDB2 is ...High-resolution fluorescence imaging using SeeDB2." Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic...strategies for combinatorial expression of fluorescent proteins in the nervous system." Nature 450.7166 (2007)...

...tagging of every protein in the C. elegans genome. Although they tested fluorescent protein fusions, this...even easier - now, one can generate a fluorescent protein fusion, transcriptional reporter, and loss-of-function... allele in just one injection step! The entire protocol takes about 2-3 weeks but requires less than eight...SEC). To make it possible fluorescently label proteins in C. elegans, they inserted the SEC into a synthetic...synthetic intron between a fluorescent protein (FP) and a 3xFLAG tag (see construct schematic below.) ...this synthetic intron, and only the fluorescent protein and 3xFLAG-tag sequences are inserted into the ...plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template and Cas9-sgRNA plasmid is...

...allows you to ask questions like: “Where does my protein of interest live within a cell,” “Does this disease...where they bind to their targets, most commonly proteins. You can then use fluorescent microscopy to look... multiplexing is that you can ask how multiple proteins relate to one another in the same sample. For ...other antibodies. But, once you do find the right protocol, multiplex IF can give you impactful and beautiful...manufacturer’s recommendations.  As you optimize your protocol, always remember to include controls to show that...experiment will have a significant impact on the protocol you use and the ultimate outcome of your assay...share some of their results and corresponding protocols. Other researchers may also have tested an antibody...

...where a fluorescent protein is inserted into one of the loop regions of any G-protein-coupled receptor (...Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein plasmids ...circularly-permuted GFP into an intracellular loop of various G-protein-coupled receptors. Image adapted from Ino et al...calcium reporter Near-infrared (NIR) fluorescent proteins promise better performance for in vivo use and...indicator using small monomeric NIR fluorescent proteins (miRFP670nano and miRFP720) and the Ca2+-sensing...structure-guided engineering of potassium binding proteins, they developed a family of fluorescent potassium...Introducing mutations and using potassium-binding protein homologues from various bacteria led to sensors...

...repair proteins fused to CGBEs The David Liu lab also experimented with adding different proteins to the... binding motif protein X-linked (RBMX) (Koblan et al., 2021). Several of these proteins had additive effects... (2) a complementary DNA sequence, and (3) a protospacer-adjacent motif (PAM) on the opposite strand. ... cytosine transversion editors. New DNA repair proteins fused to CGBEs Instead of using UNG, the Wei Leong... CBE with other BER components: the DNA repair protein rXRCC1, or the DNA binding and lyase domain of ...Mycobacterium smegmatis, termed UdgX, they found three proteins whose fusion improved the CGBE: DNA polymerase...improving C → G editing. Instead of fusing DNA repair proteins to the base editor, they added the pioneer factor...

...Vesicular stomatitis virus G glycoprotein; Broad tropism envelope protein; Pseudotyped to alter infectivity...(viral Gag-Pol): Gag is a structural precursor protein, and Pol is a polymerase. Envelope gene (may be...packaging (Gag, Pol, and Env), the isoforms of these proteins, as well as the viral LTRs, differ. As a result...plasmids, such as VSV-G (Vesicular Stomatitis Virus glycoprotein), however, may be used across both systems.  ...pseudotyping.   Figure 1: Genome of a prototypical gamma-retrovirus.    Frequently asked questions...infectivity. Packaging Gag Precursor structural protein of the retroviral particle containing Matrix, Capsid...and Nucleocapsid components. Pol Precursor protein containing Reverse Transcriptase and Integrase ...

...psychrophilic proteins. The primary problem we wanted to overcome was inefficient protein folding at low...psychrophilic proteins. As the figure below shows, E. coli expressing the chaperones protein resulted in...Following up its 2018 iGEM debut - centered around a prototypical methane biosensor - IISER Bhopal is back in ...enzymes for the host The expression of recombinant proteins for industrial or research purposes requires stable...example E. coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes are highly preferable... standard conditions. However, one of the many proteins which are not compatible with regular mesophilic... suboptimal conditions. This means recombinant proteins can aggregate or precipitate out as insoluble ...